Journal of Pathology and Translational Medicine

Case Study

Primary pulmonary epithelioid inflammatory myofibroblastic sarcoma: a rare entity and a literature review: Priyanka Singh, Aruna Nambirajan, Manish Kumar Gaur, Rahul Raj, Sunil Kumar, Prabhat Singh Malik, Deepali Jain; J Pathol Transl Med. 2022;56(4):231-237. Published online July 7, 2022; DOI: https://doi.org/10.4132/jptm.2022.05.08

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Epithelioid inflammatory myofibroblastic sarcoma (EIMS) is an aggressive subtype of inflammatory myofibroblastic tumor (IMT) harboring anaplastic lymphoma kinase (ALK) gene fusions and is associated with high risk of local recurrence and poor prognosis. Herein, we present a young, non-smoking male who presented with complaints of cough and dyspnoea and was found to harbor a large right lower lobe lung mass. Biopsy showed a high-grade epithelioid to rhabdoid tumor with ALK and desmin protein expression. The patient initially received 5 cycles of crizotinib and remained stable for 1 year; however, he then developed multiple bony metastases, for which complete surgical resection was performed. Histopathology confirmed the diagnosis of EIMS, with ALK gene rearrangement demonstrated by fluorescence in situ hybridization. Postoperatively, the patient is asymptomatic with stable metastatic disease on crizotinib and has been started on palliative radiotherapy. EIMS is a very rare subtype of IMT that needs to be included in the differential diagnosis of ALKexpressing lung malignancies in young adults.

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Inflammatory Myofibroblastic Tumor: An Updated Review
Joon Hyuk Choi
Cancers.2025; 17(8): 1327. CrossRef
Epithelioid Inflammatory Myofibroblastic Sarcoma: Case Series With a First Report of CLTC::ALK Fusion in an Aggressive Disease
Daisy Maharjan, Carina Dehner, Ali Alani, Robert Bell, Sheila Segura
Genes, Chromosomes and Cancer.2025;[Epub] CrossRef
ALK rearranged malignant mesenchymal neoplasms of thorax: therapeutically targetable ‘ALKomas’ beyond the spectrum of non-small cell lung carcinomas and thoracic inflammatory myofibroblastic tumors
Shreya Sadhu, Adarsh Barwad, Asit Ranjan Mridha, Prabhat Singh Malik, Aruna Nambirajan, Deepali Jain
Virchows Archiv.2025; 487(5): 1003. CrossRef
Mediastinal epithelioid inflammatory myofibroblastic sarcoma with the EML4‐ALK fusion: A case report and literature review
Tingyu Pan, Xinyu Sun, Xiao Wu, Futing Tang, Xianmei Zhou, Qian Wang, Shi Chen
Respirology Case Reports.2024;[Epub] CrossRef
Primary epithelioid inflammatory myofibroblastic sarcoma of the brain with EML4::ALK fusion mimicking intra-axial glioma: a case report and brief literature review
Eric Eunshik Kim, Chul-Kee Park, Koung Mi Kang, Yoonjin Kwak, Sung-Hye Park, Jae-Kyung Won
Journal of Pathology and Translational Medicine.2024; 58(3): 141. CrossRef
Epithelioid Inflammatory Myofibroblastic Sarcoma: A Report of a Rare Case
Varun Ronanki, Vaddatti Tejeswini, Inuganti Venkata Renuka, Shaik Raheema, Bakkamanthala S K Kanth
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Thoracic epithelioid inflammatory myofibroblastic sarcoma: a rare and aggressive disease with case report and literature review
Linke Yang, Pei Li, Runze Liu, Baomin Feng, Huiqing Mao, Xiaoyong Tang, Guangjian Yang
Discover Oncology.2024;[Epub] CrossRef
Epithelioid inflammatory myofibroblastic sarcoma with exceptionally long response to lorlatinib—a case report
Rafał Becht, Kajetan Kiełbowski, Justyna Żychowska, Wojciech Poncyljusz, Aleksandra Łanocha, Katarzyna Kozak, Ewa Gabrysz-Trybek, Paweł Domagała
Therapeutic Advances in Medical Oncology.2024;[Epub] CrossRef
Rare giant epithelioid inflammatory myofibroblastic sarcoma of the abdominal cavity in a child: a case report and review of the literature
Jinzhou Li, Haixing Su, Sheng Zhang, Xianyun Chen, Chongzhi Hou, Tao Cheng
Frontiers in Oncology.2024;[Epub] CrossRef
Case report: Epithelioid inflammatory myofibroblastic sarcoma treated with an ALK TKI ensartinib
Mengmeng Li, Ruyue Xing, Jiuyan Huang, Chao Shi, Chunhua Wei, Huijuan Wang
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Epithelioid Inflammatory Myofibroblastic Sarcoma With Poor Response to Crizotinib: A Case Report
Soheila Aminimoghaddam, Roghayeh Pourali
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Epithelioid inflammatory myofibroblastic sarcoma: a case report and brief literature review
Weidong Dou, Yu Guan, Tao Liu, Hang Zheng, Shuo Feng, Yingchao Wu, Xin Wang, Zhanbing Liu
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Original Article

Multistaining Optimization for Epstein-Barr Virus–Encoded RNA In Situ Hybridization and Immunohistochemistry of Formalin-Fixed Paraffin-Embedded Tissues Using an Automated Immunostainer: Jae Nam Ko, Jin Kyoung Jung, Yun Ik Park, Hwa Jeong Shin, Jooryung Huh, Sol Back, Yu Jin Kim, Jae Ho Kim, Heounjeong Go; J Pathol Transl Med. 2019;53(5):317-326. Published online August 27, 2019; DOI: https://doi.org/10.4132/jptm.2019.08.06

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Abstract PDF Supplementary Material

Background
Single staining is commonly performed for practical pathologic diagnoses. However, this method is limited in its ability to specify cellular morphology and immunophenotype and often requires consumption of limited tissue. This study aimed to describe an optimized protocol for multiple in situ hybridization (ISH) and immunohistochemistry (IHC).
Methods
The quality of multistaining was evaluated by carefully changing each step of ISH and IHC in an angioimmunoblastic T-cell lymphoma (AITL) case on a Ventana BenchMark XT automated immunostainer. The optimized protocols were also performed using another immunostainer and in 15 cases of five Epstein-Barr virus (EBV)–associated malignancies using formalin-fixed paraffin-embedded tissue.
Results
The quality of various ISHIHC staining protocols was semi-quantitatively evaluated. The best EBV-encoded RNA (EBER)-ISH/double IHC staining quality, equivalent to single staining, was obtained using the following considerations: initial EBER-ISH application, use of protease and antigen retrieval reagent (cell conditioning 1 [CC1] treatment time was minimized due to impact on tissue quality), additional baking/ deparaffinization not needed, and reduced dilution ratio and increased reaction time for primary antibody compared with single immunostaining. Furthermore, shorter second CC1 treatment time yielded better results. Multiple staining was the best quality in another immunostainer and for different types of EBV-associated malignancies when it was performed in the same manner as for the Ventana BenchMark XT as determined for AITL.
Conclusions
EBER-ISH and double IHC could be easily used in clinical practice with currently available automated immunostainers and adjustment of reagent treatment time, dilution ratio, and antibody reaction time.

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Shilah A. Bonnett, Alyssa B. Rosenbloom, Giang T. Ong, Mark Conner, Aric B.E. Rininger, Daniel Newhouse, Felicia New, Chi Q. Phan, Saskia Ilcisin, Hiromi Sato, John S. Lyssand, Gary Geiss, Joseph M. Beechem
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Review

Molecular Testing of Lymphoproliferative Disorders: Current Status and Perspectives: Yoon Kyung Jeon, Sun Och Yoon, Jin Ho Paik, Young A Kim, Bong Kyung Shin, Hyun-Jung Kim, Hee Jeong Cha, Ji Eun Kim, Jooryung Huh, Young-Hyeh Ko; J Pathol Transl Med. 2017;51(3):224-241. Published online May 10, 2017; DOI: https://doi.org/10.4132/jptm.2017.04.09

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Abstract PDF

Molecular pathologic testing plays an important role for the diagnosis, prognostication and decision of treatment strategy in lymphoproliferative disease. Here, we briefly review the molecular tests currently used for lymphoproliferative disease and those which will be implicated in clinical practice in the near future. Specifically, this guideline addresses the clonality test for B- and T-cell proliferative lesions, molecular cytogenetic tests for malignant lymphoma, determination of cell-of-origin in diffuse large B-cell lymphoma, and molecular genetic alterations incorporated in the 2016 revision of the World Health Organization classification of lymphoid neoplasms. Finally, a new perspective on the next-generation sequencing for diagnostic, prognostic, and therapeutic purpose in malignant lymphoma will be summarized.

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Jieun Kim, Ho Hyun Song, Soobin Chae, GeonWoo Choi, Jeong Won Shin
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Case Reports

Primary Endometrial Squamous Cell Carcinoma: A Case Report and Review of Relevant Literature on Korean Women: Sung Jong Lee, Hyun Joo Choi; Korean J Pathol. 2012;46(4):395-398. Published online August 23, 2012; DOI: https://doi.org/10.4132/KoreanJPathol.2012.46.4.395

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Abstract PDF

Primary endometrial squamous cell carcinoma (PESCC) is an extremely rare tumor with unclear pathogenesis. A 54-year-old postmenopausal woman presented with a 6-month history of vaginal bleeding. The patient was provisionally diagnosed with uterine submucosal leiomyoma. This was followed by total hysterectomy with a bilateral salpingo-oophorectomy under the laparoscopic guidance. Histopathologically, the tumor was PESCC which was accompanied by a lack of the tumor in the uterine cervix. The tumor showed positive immunoreactivity for p^16INK4a. But there was no evidence of human papillomavirus (HPV) on in situ hybridization and HPV DNA chip analysis. We also present a review of the relevant literature on Korean women.

Citations

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Primary endometrial squamous cell carcinoma (PESCC): Review of the literature and case report
Kuang-Han Liu, Chia-Chin Tsai, Krystal Baysan Lin, Pei-Shen Huang
Taiwanese Journal of Obstetrics and Gynecology.2025; 64(1): 159. CrossRef
Primary squamous cell carcinoma of the uterine wall associated with adenomyosis: A rare case report and literature review
Yang Li, Lanlan Xuan, Qiucheng Wang
International Journal of Gynecology & Obstetrics.2025;[Epub] CrossRef
p16 Block Type Overexpression, p53 Wild Type Reactivity, and Cervical Involvement do not Always Exclude the Diagnosis of Primary Endometrial Squamous Cell Carcinoma (PESCC)
Daniela Fanni, Clara Gerosa, Michele Peiretti, Valerio Mais, Elena Massa, Stefano Angioni, Gavino Faa
International Journal of Gynecological Pathology.2024; 43(2): 200. CrossRef
Pathogenetic characteristics of endometrioid adenocarcinoma of uterus at present stage
T. I. Moiseenko, S. V. Shatalova, E. M. Nepomnyashchaya, V. A. Bandovkina, M. L. Adamyan
Medical alphabet.2024; (36): 35. CrossRef
Case report: Clinicopathological characteristic of two cases of primary endometrial squamous cell carcinoma and review of the literature
Hui-Bin Zhang, Li-Hua Lin, Qiu-Ping Lin, Yuan-Qing Lin, Dan Luo, Shu-Xia Xu
Frontiers in Oncology.2024;[Epub] CrossRef
Treatment of primary squamous cell carcinoma of the endometrium and review of previous literature: A case report
Liyun Song, Qi Wu, Suning Bai, Ren Xu, Xiaona Wang, Yanyan Yang
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Prevalence of human papilloma virus and Chlamydia trachomatis in endometrial and cervical carcinoma: a comparative study in North Indian women
Heena Gautam, Sumita Mehta, Nidhi Nayar, Neha Kumar, Syed Akhtar Husain, Mausumi Bharadwaj
Systems Biology in Reproductive Medicine.2023; 69(6): 399. CrossRef
PAX8 Positivity, Abnormal p53 Expression, and p16 Negativity in a Primary Endometrial Squamous Cell Carcinoma: A Case Report and Review of the Literature
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International Journal of Gynecological Pathology.2022; 41(4): 431. CrossRef
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Sanda Rajhvajn, Ana Barišić, Lada Škopljanac‐Mačina, Danijela Jurič, Vesna Mahovlić
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Primary Pulmonary Myxoid Liposarcoma with Translocation t(12;16)(q13;p11) in a Young Female Patient: A Brief Case Report: Choonhee Son, Phil Jo Choi, Mee Sook Roh; Korean J Pathol. 2012;46(4):392-394. Published online August 23, 2012; DOI: https://doi.org/10.4132/KoreanJPathol.2012.46.4.392

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Abstract PDF

Primary liposarcoma of the lung is an extremely rare disease. To date, only 14 cases have been reported in the literature. We experienced a case of myxoid liposarcoma of the lung treated by surgery. The tumor was well-defined, solid, lobulated mass measuring 3.5×2 cm, involving the bronchus of the left lower lobe. Microscopically, myxoid liposarcoma was identified. The fluorescence in situ hybridization confirmed the presence of a reciprocal translocation involving DNA damage-inducible transcript 3 (DDIT3) and fused in sarcoma (FUS) genes. The patient is still alive with no recurrence or metastasis at the time of writing this report (on 20 months postoperatively). To our knowledge, this is the first cytogenetic case report of pulmonary myxoid liposarcoma.

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Elisa M. Wächtershäuser, Gabriele Köhler, Verena Böhmer, Alexander Marx, Achim Hellinger
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Primary intrathoracic liposarcomas: A clinicopathologic and molecular study of 43 cases in one of the largest medical centers of China
You Xie, Wenyi Jing, Wei Zhao, Ran Peng, Min Chen, Ting Lan, Heng Peng, Xin He, Huijiao Chen, Zhang Zhang, Hongying Zhang
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Primary Pulmonary Liposarcoma with Pancreatic Metastasis: A Rarest of Rare Intrathoracic Malignancy
Anirban Halder, Rituparna Biswas, Sujit Shukla, Nisha Rana, Vikas Yadav, Jaspreet Kaur
Indian Journal of Medical and Paediatric Oncology.2020; 41(04): 605. CrossRef
Primary dedifferentiated liposarcoma of the lung with rhabdomyoblastic and chrondroblastic differentiation
Anthony Longano, Alexandra DuGuesclin, Catherine Mitchell
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Original Articles

Diagnostic Value of MDM2 and DDIT3 Fluorescence In Situ Hybridization in Liposarcoma Classification: A Single-Institution Experience: Junhun Cho, Seung Eun Lee, Yoon-La Choi; Korean J Pathol. 2012;46(2):115-122. Published online April 25, 2012; DOI: https://doi.org/10.4132/KoreanJPathol.2012.46.2.115

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Abstract PDF

Background

The amplification of murine double minutes (MDM2) is the primary feature of well-differentiated liposarcomas (WDLPS) and dedifferentiated liposarcomas (DDLPS), while DDIT3 rearrangement is the main one of myxoid liposarcomas (MLPS). Our aim was to evaluate the added value of MDM2 amplification and DDIT3 rearrangement in making a diagnosis and classifying lipogenic tumors.

Methods

Eighty-two cases of liposarcoma and 60 lipomas diagnosed between 1995 and 2010 were analysed for MDM2 amplification and DDIT3 rearrangement using a fluorescence in situ hybridization (FISH). The subtypes of liposarcoma were reclassified according to the molecular results, whose results were reviewed with an analysis of the relevant histologic and immunohistochemical findings.

Results

One case of lipoma (1.67%) was reclassified as a WDLPS. Of the liposarcomas, 13.4% (16/82) were reclassified after the molecular testing. Five cases of MLPS were reclassified as four cases of DDLPS and one case of myxoid lipoma. Two cases of WDLPS were reclassified as one case of spindle cell lipoma and another case of myxofibrosarcoma. Four cases of DDLPS were reclassified as two cases of leiomyosarcoma, one case of angiomyolipoma and another case of fibroinflammatory lesion. Of the six cases of pleomorphic liposarcoma, five were reclassified as DDLPS.

Conclusions

In our series, a critical revision of diagnosis was found at a rate of 3.5% (5/142) after a review of the lipomatous lesions. The uses of molecular testing by MDM2 and DDIT3 FISH were valuable to make an accurate subtyping of liposarcomas as well as to differentiate WDLPS from benign lipomatous tumor.

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Myxoid liposarcoma with nuclear pleomorphism: a clinicopathological and molecular study
Naoki Kojima, Takashi Kubo, Taisuke Mori, Kaishi Satomi, Yuko Matsushita, Shintaro Iwata, Yasushi Yatabe, Koichi Ichimura, Akira Kawai, Hitoshi Ichikawa, Akihiko Yoshida
Virchows Archiv.2024; 484(1): 71. CrossRef
FISH Diagnostic Assessment of MDM2 Amplification in Liposarcoma: Potential Pitfalls and Troubleshooting Recommendations
Alessandro Gambella, Luca Bertero, Milena Rondón-Lagos, Ludovica Verdun Di Cantogno, Nelson Rangel, Chiara Pitino, Alessia Andrea Ricci, Luca Mangherini, Isabella Castellano, Paola Cassoni
International Journal of Molecular Sciences.2023; 24(2): 1342. CrossRef
Expression of CTAG1B clone EPR13780 versus DDIT3 gene rearrangement distinguishes myxoid liposarcoma from its mimics with detection of novel DDIT3 gene copy number variations
Marwa M. Abdelaziz, Hanan Y. Tayel, Amany Abdel-Bary, Omnia M. Badawy
Journal of Histotechnology.2022; 45(2): 56. CrossRef
Musculoskeletal Tumors
Amit Singla, David S. Geller
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Vulvar Myxoid Liposarcoma, an Extremely Rare Diagnosis: A Case Report and Review of Literature
Ligia Redroban, Nelson Montalvo
International Journal of Gynecological Pathology.2019; 38(1): 17. CrossRef
Molecular updates in adipocytic neoplasms✰
Elizabeth G. Demicco
Seminars in Diagnostic Pathology.2019; 36(2): 85. CrossRef
Application of MDM2 Fluorescence In Situ Hybridization and Immunohistochemistry in Distinguishing Dedifferentiated Liposarcoma From Other High-grade Sarcomas
Min Jeong Song, Kyung-Ja Cho, Jong-Seok Lee, Joon Seon Song
Applied Immunohistochemistry & Molecular Morphology.2017; 25(10): 712. CrossRef
FluorescenceIn SituHybridization forMDM2Amplification as a Routine Ancillary Diagnostic Tool for Suspected Well-Differentiated and Dedifferentiated Liposarcomas: Experience at a Tertiary Center
Khin Thway, Jayson Wang, John Swansbury, Toon Min, Cyril Fisher
Sarcoma.2015; 2015: 1. CrossRef
Complete response of a recurrent-metastatic liposarcoma with dedifferentiated histological features following the administration of trabectedin and review of literature
Tulay Kus, Gokmen Aktas, Mehmet Emin Kalender, Ediz Tutar, Esra Ulker, Celaletdin Camci
Journal of Cancer Research and Therapeutics.2015; 11(4): 974. CrossRef

Prognostic Significance of Amplification of the c-MYC Gene in Surgically Treated Stage IB-IIB Cervical Cancer.: Tae Jung Kim, Ahwon Lee, Sung Jong Lee, Won Chul Lee, Yeong Jin Choi, Kyo Young Lee, Chang Suk Kang; Korean J Pathol. 2011;45(6):596-603.; DOI: https://doi.org/10.4132/KoreanJPathol.2011.45.6.596

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Abstract PDF

BACKGROUND
Mutations of c-MYC have been described in cervical cancer. However, association between c-MYC gene status and its prognostic significance have not been clarified.
METHODS
Tissue microarray sections from 144 patients with stage IB-IIB cervical cancer treated by radical hysterectomy were analyzed by fluorescence in situ hybridization using a region-specific probe for c-MYC and a centromere-specific probe for chromosome 8.
RESULTS
Seventy five percent (108/144) of c-MYC gain and 6.9% (10/144) of c-MYC gene amplification were observed. c-MYC gene alteration was more frequently observed in squamous cell carcinoma than adenocarcinoma or adenosquamous carcinoma and were associated with low Ki67 labeling index (p=0.013). c-MYC amplification was not associated with clinicopathologic parameters except absence of bcl2 expression (p=0.048). Survival analysis revealed that patients with c-MYC amplification were significantly associated with higher risk of disease recurrence (p=0.007) and cancer related death (p=0.020). However, c-MYC gain was not associated with unfavorable outcome. Multivariate analysis proved c-MYC amplification as independent prognostic factors of shorter disease free survival and cancer-related death (p=0.028 and p=0.025, respectively).
CONCLUSIONS
c-MYC amplification, not gain, is an independent prognostic marker for shorter disease free and cancer specific survival in cervical cancer treated by radical hysterectomy.

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A Rare Case of Cutaneous Plasmacytosis in a Korean Male
Corey Georgesen, Meenal Kheterpal, Melissa Pulitzer
Case Reports in Pathology.2017; 2017: 1. CrossRef

Evaluation of the HPV ISH Assay in Cervical Cancer.: Jung Uee Lee, Jung Ha Shin, Jong Ok Kim, Yeong Jin Choi, Kyo Young Lee, Jong Sup Park, Won Chul Lee, Ahwon Lee; Korean J Pathol. 2010;44(5):513-520.; DOI: https://doi.org/10.4132/KoreanJPathol.2010.44.5.513

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Abstract PDF

BACKGROUND
Human papillomavirus (HPV) infection can be detected by in situ hybridization (ISH), in which a punctate signal pattern indicates integrated HPV DNA and a diffuse pattern denotes the presence of episomal viral DNA. This study was conducted to evaluate the usefulness of an HPV ISH assay for invasive cervical cancer.
METHODS
The HPV ISH assay for high-risk HPV and immunohistochemical staining for p16(INK4a), p53, bcl-2, and Ki-67 were performed in a tissue microarray of 279 cervical cancers.
RESULTS
High-risk HPV ISH was positive in 194 (69.5%) of the samples. Punctate, diffuse, and mixed signal patterns were observed in 157 (56.3%), one (0.4%), and 36 cases (12.9%), respectively. Positive results in high-risk HPV ISH were associated with p16 and bcl-2 expression (p = 0.01 and p < 0.01, respectively). According to a Cox regression analysis, HPV infection and its surrogate immunohistochemical markers such as p16, bcl-2, and Ki-67 were not independent prognostic factors, but stage and grade were independent prognostic factors.
CONCLUSIONS
Our results confirm that an HPV ISH assay is reasonably sensitive for HPV infection and that it might be useful to identify integrated HPV DNA in formalin-fixed and paraffin-embedded specimens. Further study encompassing HPV type, E2/E6 ratio, and therapeutic modality is necessary to understand the clinical meaning of HPV status in cervical cancer.

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Shaikhali M Barodawala, Kirti Chadha, Vikas Kavishwar, Anuradha Murthy, Shamma Shetye
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Prognostic Significance of Amplification of thec-MYCGene in Surgically Treated Stage IB-IIB Cervical Cancer
Tae-Jung Kim, Ahwon Lee, Sung-Jong Lee, Won-Chul Lee, Yeong-Jin Choi, Kyo-Young Lee, Chang Suk Kang
The Korean Journal of Pathology.2011; 45(6): 596. CrossRef

HER2 Status in Gastric Adenocarcinomas Assessed by Immunohistochemistry, Automated Silver-Enhanced In Situ Hybridization and Fluorescence In Situ Hybridization.: Aeri Kim, Jung Min Bae, Se Won Kim, Mi Jin Gu, Young Kyung Bae; Korean J Pathol. 2010;44(5):493-501.; DOI: https://doi.org/10.4132/KoreanJPathol.2010.44.5.493

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Abstract PDF

BACKGROUND
Recently, many studies have focused on human epidermal growth factor receptor 2 (HER2) status in gastric cancer due to HER2-targeted therapy using trastuzumab. We investigated HER2 overexpression and amplification and their concordance rate in Korean gastric adenocarcinomas.
METHODS
Tissue microarrays were constructed with 232 gastric adenocarcinoma samples. We performed immunohistochemistry (IHC), silver-enhanced in situ hybridization (SISH) and fluorescence in situ hybridization (FISH) for HER2.
RESULTS
IHC was negative in 94.8% (218/232), equivocal in 1.7% (4/232) and positive in 3.5% (8/232) of cases. HER2 protein expression was heterogeneous in 75% (9/12) of IHC 2+/3+ cancers. Gene amplification was observed in 6.5% (15/230) by SISH and the same 15 cases were also FISH-positive. We observed HER2 amplification in 1.4%, 27.3%, 25%, and 100% of IHC 0, 1+, 2+, and 3+ gastric adenocarcinomas, respectively. The concordance rate between IHC and SISH results was 95.7%.
CONCLUSIONS
HER2 overexpression and amplification were less frequent in gastric adenocarcinomas than breast carcinomas. Compared to breast carcinoma, (1) there may be IHC-negative but gene amplification-positive cases for HER2 and (2) frequent intratumoral heterogeneity of IHC for HER2 in gastric adenocarcinomas.

Citations

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Epidemiologic Study of Human Epidermal Growth Factor Receptor 2 Expression in Advanced/Metastatic Gastric Cancer: an Assessment of Human Epidermal Growth Factor Receptor 2 Status in Tumor Tissue Samples of Gastric and Gastro-Esophageal Junction Cancer
Kyung Won Seo, Taeyong Jeon, Sewon Kim, Sung Soo Kim, Kwanghee Kim, Byoung-Jo Suh, Sunhwi Hwang, SeongHee Choi, Seungwan Ryu, Jae Seok Min, Young-Joon Lee, Ye Seob Jee, Hyeondong Chae, Doo Hyun Yang, Sang Ho Lee
Journal of Gastric Cancer.2017; 17(1): 52. CrossRef
Synopsis on Clinical Practice Guideline of Gastric Cancer in Korea: An Evidence-Based Approach
Jun Haeng Lee, Jae G. Kim, Hye-Kyung Jung, Jung Hoon Kim, Woo Kyoung Jeong, Tae Joo Jeon, Joon Mee Kim, Young Il Kim, Keun Won Ryu, Seong-Ho Kong, Hyoung Il Kim, Hwoon-Yong Jung, Yong Sik Kim, Dae Young Zang, Jae Yong Cho, Joon Oh Park, Do Hoon Lim, Eun S
The Korean Journal of Gastroenterology.2014; 63(2): 66. CrossRef
Clinical Practice Guidelines for Gastric Cancer in Korea: An Evidence-Based Approach
Jun Haeng Lee, Jae G. Kim, Hye-Kyung Jung, Jung Hoon Kim, Woo Kyoung Jeong, Tae Joo Jeon, Joon Mee Kim, Young Il Kim, Keun Won Ryu, Seong-Ho Kong, Hyoung-Il Kim, Hwoon-Yong Jung, Yong Sik Kim, Dae Young Zang, Jae Yong Cho, Joon Oh Park, Do Hoon Lim, Eun S
Journal of Gastric Cancer.2014; 14(2): 87. CrossRef

Automated Silver-enhanced In Situ Hybridization for Evaluation of HER2 Gene Status in Breast Carcinoma: Comparison with Fluorescence In Situ Hybridization and Immunohistochemistry.: Woo Jung Sung, Seok Ju Park, Mi Jin Gu, Young Kyung Bae; Korean J Pathol. 2010;44(1):28-34.; DOI: https://doi.org/10.4132/KoreanJPathol.2010.44.1.28

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Abstract PDF

BACKGROUND
The human epidermal growth factor receptor 2 (HER2) is amplified in 20-25% of breast cancers. HER2 overexpression or amplification is associated with a worse clinical outcome and it can predict the benefit from anthracycline and anti-HER2 therapies. The HER2 status has usually been assessed by immunohistochemistry (IHC) or fluorescence in situ hybridization (FISH) in clinical samples. A new silver-enhanced in situ hybridization (SISH) technique was recently introduced. Therefore we evaluated the usefulness of SISH for detecting HER2 amplification.
METHODS
Tissue microarrays (TMAs) were constructed with 144 invasive breast cancer tissue samples. We performed IHC, FISH and SISH for HER2 on the tissue sections from the TMAs and we interpreted the results according to the American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) guidelines. The concordant rates between the two different tests were calculated.
RESULTS
HER2 was overexpressed and amplified in 16.9%, 16.9%, and 18% of the cases by IHC, FISH and SISH, respectively. The concordant rates between IHC and FISH, IHC and SISH, and FISH and SISH were 95.1%, 95.7%, and 97.8%, respectively.
CONCLUSIONS
SISH can be an alternative test for evaluating HER2 amplification because the 97.8% concordance with FISH satisfies the ASCO/CAP requirement of > 95% concordance with an alternative validated method.

Citations

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HER2 assessment by silverin situhybridization: where are we now?
Francesca Sanguedolce, Pantaleo Bufo
Expert Review of Molecular Diagnostics.2015; 15(3): 385. CrossRef
Diagnostic Effectiveness of PCR-based Tests DetectingBRAFMutation for Treating Malignant Melanoma: A Systematic Review
Hae-Won Shin, Ryeo-Jin Ko, Min Lee, Hee-Young Bang, Kye-Chul Kwon, Jong-Woo Park, Sun-Hoe Koo
Laboratory Medicine Online.2014; 4(4): 203. CrossRef
Effectiveness of Silver-enhanced In Situ Hybridization for Evaluating HER2 Gene Status in Invasive Breast Carcinoma: A Comparative Study
Youngseok Lee, Youngjoon Ryu, Hoiseon Jeong, Hyeyoon Chang, Younghye Kim, Aeree Kim
Archives of Medical Research.2012; 43(2): 139. CrossRef
HER2 Status by Standardized Immunohistochemistry and Silver-EnhancedIn SituHybridization in Korean Breast Cancer
Young Kyung Bae, Gyungyub Gong, Jun Kang, Ahwon Lee, Eun Yoon Cho, Ji Shin Lee, Kwang-Sun Suh, Dong Wha Lee, Woo Hee Jung
Journal of Breast Cancer.2012; 15(4): 381. CrossRef
Multiplication of Chromosome 17 Centromere Is Associated with Prognosis in Patients with Invasive Breast Cancers Exhibiting NormalHER2andTOP2AStatus
Aeri Kim, Hyung Chan Shin, Young Kyung Bae, Min Kyoung Kim, Su Hwan Kang, Soo Jung Lee, Eun Hee Lee
Journal of Breast Cancer.2012; 15(1): 24. CrossRef
HER2-Positive Breast Carcinomas with Co-amplification or Gain of Chromosome 17 Centromere Locus: Report of Three Cases and an Impact on HER2 Testing
Hyeong Chan Shin, Young Kyung Bae, Aeri Kim, Seok Ju Park
The Korean Journal of Pathology.2011; 45(6): 665. CrossRef
The Effectiveness of SilverIn SituHybridization in Patients with Breast Cancer: A Systematic Review
Sunyoung Jang, Seon-Heui Lee, Soojin Kim, You-Kyoung Lee, Young-Hyuck Im, Wonshik Han, Hee-Sook Park
Journal of Breast Cancer.2011; 14(Suppl 1): S1. CrossRef
Clinical and health economic outcomes of alternative HER2 test strategies for guiding adjuvant trastuzumab therapy
James A Lee, Megan Shaheen, Thomas Walke, Matt Daly
Expert Review of Pharmacoeconomics & Outcomes Research.2011; 11(3): 325. CrossRef
HER2 expression in breast cancer: Comparisons of fluorescence in situ hybridization and silver in situ hybridization, and immunohistochemical staining using monoclonal antibody and polyclonal antibody
Jung Sik Jang, Eun Jeong Jang, Ji‐Young Park
Basic and Applied Pathology.2010; 3(4): 115. CrossRef
HER2Status in Gastric Adenocarcinomas Assessed by Immunohistochemistry, Automated Silver-EnhancedIn SituHybridization and FluorescenceIn SituHybridization
Aeri Kim, Jung Min Bae, Se Won Kim, Mi Jin Gu, Young Kyung Bae
The Korean Journal of Pathology.2010; 44(5): 493. CrossRef

The Prognostic Subgroups as Defined by the Patterns of Epstein-Barr Virus Infection in Patients with Hodgkin Lymphoma.: Ji Hyeon Roh, Seok Jin Kim, Won Seog Kim, Young Hyeh Ko; Korean J Pathol. 2010;44(1):22-27.; DOI: https://doi.org/10.4132/KoreanJPathol.2010.44.1.22

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Abstract PDF: BACKGROUND
The purpose of this study was to investigate the prognostic significance of the Epstein-Barr virus (EBV) infected non-neoplastic lymphocytes in patients with Hodgkin lymphoma (HL).
METHODS
Seventy-seven cases of HL were evaluated by immunohistochemical analysis and EBV-encoded RNA in situ hybridization. The cases were divided into three groups according to the EBV status. EBV was negative in 48 cases (group 1), EBV was located in the Hodgkin/Reed-Sternberg (HRS) cells, but not in the non-neoplastic lymphocytes in 20 cases (26%) (group 2) and EBV was located in both the HRS cells and the non-neoplastic lymphocytes in 9 cases (12%) (group 3).
RESULTS
The groups differed with respect to the age distribution, the clinical presentation and the prognosis. The median ages were 30 (group 1), 47.5 (group 2) and 23 years (group 3) (p = 0.011). B symptoms (p = 0.007) and the histologic subtype of mixed cellularity classical HL (p = 0.001) were more common in the EBV-positive patients than in their EBV-negative counterparts. Two patients from group 3 had associated chronic EBV infection syndrome. The five-year survival rate was 97.56% in group 1, 75.76% in group 2 and 100% in group 3 (p = 0.0178).
CONCLUSIONS
HL with EBV located in both the HRS cells and the non-neoplastic lymphocytes is a distinct prognostic subgroup that has better survival than the HL with EBV located in only the HRS cells.

Case Report

Extramedullary B Lymphoblastic Crisis of CML, Presenting as a Leptomeningeal Tumor: A Case Report.: Hoiseon Jeong, Bongkyung Shin, Sook Young Bae, Sangho Lee, Youngjune Ryu, Hankyeom Kim, Insun Kim; Korean J Pathol. 2009;43(5):482-488.; DOI: https://doi.org/10.4132/KoreanJPathol.2009.43.5.482

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Abstract PDF: We report here on a rare case of a patient who presented with an extramedullary B lymphoblastic crisis as an initial manifestation of chronic myelogenous leukemia (CML). A 71-year-old man visited the emergency room due to suddenly developed dysarthria and right side weakness. Emergency craniotomy was done under the presumptive diagnosis of subdural hemorrhage. During the operation, a poorly demarcated firm mass was identified in the leptomeningeal space. Microscopically, the majority of the tumor was composed of monotonous immature cells with blast morphology, and these cells were immunoreactive for TdT, CD34, CD10 and CD20, indicating the precursor B-cell phenotype. The peripheral area of the tumor consisted of myeloid cells in various stages of maturation, and these cells were reactive for myeloperoxidase, chloroacetate esterase, CD43 and CD15. FISH analysis using the LSI bcr-abl dual color probe showed gene fusion signals in both the B-lymphoblasts and myeloid cells. The peripheral blood and bone marrow findings were consistent with CML with no evidence of a blast crisis. Cytogenetic study of the bone marrow demonstrated the 46, XY, t(9;22)(q34;q11) chromosome. A diagnosis of extramedullary B lymphoblastic blast crisis in a patient with Philadelphia chromosome-positive CML was made. Despite treatment, the patient died 3 months after he was diagnosed.

Original Articles

The EGFR Protein Expression and the Gene Copy Number Changes in Renal Cell Carcinomas.: Sangho Lee, Jungsuk An, Aeree Kim, Young Sik Kim, Insun Kim; Korean J Pathol. 2009;43(5):413-419.; DOI: https://doi.org/10.4132/KoreanJPathol.2009.43.5.413

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Abstract PDF

BACKGROUND
The epidermal growth factor receptor (EGFR) is known to be involved in many tumor promoting activities. EGFR inhibition has been tried as a therapeutic modality in many human malignancies.
METHODS
The expression of EGFR protein and the gene copy number changes were studied in 135 clear cell carcinomas and 16 papillary renal cell carcinomas (RCCs), and these tumors were diagnosed between 1995 and 1997.
RESULTS
An EGFR protein expression (2+ and 3+) was found in 54.1% of the clear cell RCCs and in 43.8% of the papillary RCCs. In the clear cell RCCs, its expression was associated with male gender, the tumor size (> or =4 cm) and high T stages (T2 and T3), with statistical significance. Trisomy and polysomy of the EGFR gene were found in 27 (25.7%) and 40 (38.1%) of 105 clear cell RCCs, respectively. Trisomy and polysomy were correlated with an EGFR protein expression and a high clinical T stage, with statistical significance. Among 15 papillary RCCs, 13 tumors showed trisomy (86.7%) and one showed polysomy (6.7%). Amplification was not found in both the clear cell and papillary type RCCs.
CONCLUSIONS
A considerable numbers of RCCs showed an overexpression of EGFR protein and increased EGFR gene copy numbers, yet the clinical significance of conducting a FISH study in RCC patients seems to be limited.

Citations

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EGFR protein overexpression correlates with chromosome 7 polysomy and poor prognostic parameters in clear cell renal cell carcinoma
Gordana Đorđević, Koviljka Matušan Ilijaš, Ita Hadžisejdić, Anton Maričić, Blaženka Grahovac, Nives Jonjić
Journal of Biomedical Science.2012;[Epub] CrossRef

Telomerase mRNA Expression by In Situ Hybridization in Premalignant Lesions and Carcinomas of the Breast.: Young Kyung Bae, Dong Sug Kim, Soo Jung Lee, Koing Bo Kwun; Korean J Pathol. 2001;35(1):53-59.

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Abstract PDF: BACKGROUND
Telomerase is a ribonucleoprotein, DNA polymerase that synthesizes telomere repeats onto chromosomal ends and maintains telomere length. Telomerase activity has been detected in a broad range of human malignant neoplasms, but not in normal somatic cells. So, activation of telomerase may represent an essential step in the malignant transformation of cells. However, the expression of telomerase in premalignant lesions remains relatively unexplored. This study was conducted to investigate the reactivation of telomerase in the carcinogenesis of human breast tissue.
METHODS
In situ hybridization for the telomerase RNA component (human telomerase mRNA; hTR) was used in a normal breast tissue (n=41), florid ductal hyperplasia (FDH) (n=10), atypical ductal hyperplasia (ADH) (n=3), ductal carcinoma in situ (DCIS) (n=44) and invasive carcinoma (n=33). hTR expression in relation to p53 status and the pathologic parameters in breast cancer was also studied.
RESULTS
Expression of hTR was demonstrated in 13 samples (31.7%) of normal breast tissues, 4 (40%) of FDH, 3 (100%) of ADH, 42 (95.5%) of DCIS, and 33 (100%) of invasive carcinoma. The rate of hTR expression of ADH was significantly different from that of FDH (p<0.05), and there were no differences in hTR expression rates among ADH, DCIS and invasive carcinomas. There was no correlation between hTR expression and nuclear grade, tumor size, and p53 status in invasive carcinomas.
CONCLUSION
These results suggest that telomerase activation may be an early event and an essential step in the carcinogenesis of human breast tissue, and that telomerase has no correlations with p53 status and prognostic parameters.

Improved Technique of Digoxigenin Labeled RNA in situ Hybridization.: Suk Keun Lee, Yeon Sook Kim, In Sun Song, Sang Shin Lee, Young Jun Lee, Woo Ho Kim, Je Geun Chi; Korean J Pathol. 2001;35(2):98-110.

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Abstract PDF: BACKGROUND
A practical RNA in situ hybridization method using digoxigenin labeled RNA probes is described in order to evaluate the technical difficulties and problems in RNA in situ hybridization.
METHODS
The paraffin sections, routinely processed in the Pathology Laboratory, were tested for the possibility of RNA in situ hybridization instead of the RNase free paraffin sections, fixed in 4% paraformaldehyde and prepared using RNase protection procedures.
RESULTS
Most of the paraffin sections, fixed in 10% neutral formalin solution in fresh condition, showed relatively good reaction of RNA in situ hybridization, although the necrotic tissue and autopsy specimens showed poor reaction of RNA in situ hybridization. A refixation procedure using a 4% paraformaldehyde solution was evaluated for optimal expression of mRNA in the paraffin sections.
CONCLUSION
The treatment of 4% paraformaldehyde before the treatment of proteinase K showed better in situ hybridization than did the treatment of 4% paraformaldehyde after the treatment of proteinase K. Also a new Polymerase Chain Reaction (PCR)-based method of RNA probe production showed consistently good results.

Detection of Human Papillomavirus in Lesions of Uterine Cervix Immunohistochemistry and in situ Hybridization.: Chang Soo Park, Jong Hee Nam, Jae Hyuk Lee, Jong Soon Kim, Seung Jin Oh; Korean J Pathol. 1997;31(4):289-297.

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Abstract PDF: To evaluate the detection of HPV DNA according to subtype of lesions of uterine cervix and its clinical applicability, in situ hybridization (ISH) and immunohistochemistry for HPV were performed in 189 cases of uterine cervical lesion, including 23 cases of low grade squamous intraepithelial lesion (SIL), 115 cases of high grade SIL and 51 cases of invasive carcinoma. Positive immunostaining, brown precipitate, was mainly noted in the nucleus of koilocytes in the superficial and intermediate layer. Positivity of immunostaining was 21.7% in low grade SIL, 13.0% in high grade SIL and 9.8% in invasive carcinoma. Positive reaction in ISH, red precipitate, was noted in the nucleus of not only koilocytes but also non-koilocytes in the superficial and intermediate layer, and dot precipitate was rarely identified in the nest of squamous cell carcinoma. Based on HPV subtype, 6/11 was 21.7% in low grade SIL, 16/18 was 32.2% and 39.2% in high grade SIL and invasive carcinoma, respectively. With regard to their associated HPV types, low grade SILs were heterogeneous and high grade SILs and invasive carcinomas were related with the high oncogenic risk group only. The correlation of HPV subtypes with panHPV was 91.3% in low grade SIL, 91.3% in high grade SIL and 98.0% in invasive carcinoma. These results suggest that detection of HPV infection by ISH may be a more useful method than immunohistochemistry and application of the HPV subtype probe with the panHPV probe could improve the sensitivity of ISH.

Clinicopathologic Analysis of Epidermal Growth Factor Receptor Status in Non-small Cell Lung Cancer: Protein Expression, Gene Amplification and Survival Analysis.: Seungkoo Lee, Jene Choi, Se Jin Jang; Korean J Pathol. 2007;41(6):387-392.

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Abstract PDF: BACKGROUND
Abnormal over-expression or gene amplification of epidermal growth factor receptor (EGFR) is important in the prognosis of non-small cell lung cancer (NSCLC). We investigated the frequency of EGFR protein expression and gene amplification, and the correlation between EGFR status and survival in NSCLC.
METHODS
We examined 360 cases of microarrayed NSCLC tissues for the EGFR protein expression and EGFR gene amplification using immunohistochemistry and fluorescent in situ hybridization.
RESULTS
EGFR protein expression and EGFR gene amplification occurred in 110 cases (30.6%) and 24 cases (6.7%), respectively. EGFR protein expression and gene amplification were more frequent in squamous cell carcinoma than in adenocarcinoma. Differences in EGFR protein expression did not dramatically affect survival curves (p=0.740), but differences in gene amplification did (p<0.05): EGFR gene amplification was associated with a lower 5-year survival rate.
CONCLUSION
EGFR protein expression and gene amplification showed moderate correlation with each other. EGFR gene amplification predicted a poor prognosis, whereas EGFR protein expression did not.

Detection of Human Papillomavirus DNA by In Situ Hybridization using Biotinylated DNA Probes in Cervical Intraepithelial Neoplasias and Squamous Cell Carcinomas.: Sang Sook Lee, Ki Kwon Kim, Chai Hong Chung, Seung Won Jin, U Ik Sohn; Korean J Pathol. 1990;24(1):16-26.

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Abstract PDF: The authors examined 9 condylomas, 26 cervical intraepithelial neoplasms(CIN) and 22 invasive squamous cell carcinomas for the presence of human papillomavirus(HPV) DNA sequences by DNA-DNA in situ hybridization. In situ hybridization revealed target HPV DNA sequences mostly in the nuclei of the superficial cells from epithelium which contained either maturation or koilocytotic atypias. With the use of biotinylated HPV DNA probes 6/11, 16/18 and 31/33/35, 42 of the 57(73.7%) were positive with HPV-6/11, 23 with HPV-16/18, 32 with HPV-31/33/35 and 18 with two or more mixed probes. HPV-31/33/35 was wht most prevalent in CIN and invasive squamous cell carcinomas, follwed by HPV-16/18. The incidence of HPV DNA increased from 66.7% to 86.4% with increasing severity of the lesions from condylomas to invasive squamous cell carcinomas. Flat condyloma was most freuently accompanied by CIN.

In Situ Detection of mRNA and RNA Component of Human Telomerase in Proliferative Lesions of the Stomach.: Mi Sook Kim, Sang Woo Juhng; Korean J Pathol. 2001;35(4):299-305.

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Abstract PDF: BACKGROUND
Proliferative lesions of the stomach were investigated by in situ hybridization using RNA probes for telomerase components and compared with the results by TRAP (telomeric repeat amplification protocol) assay.
METHODS
RNA probes for hTR (human telomerase RNA component) and hTERT (mRNA coding for a catalytic subunit of human telomerase) were made by cloning and in vitro transcription. The probes were applied for in situ hybridization in 23 cases of adenocarcinoma of the intestinal type and adjacent dysplasia, and in the normal and metaplastic mucosa of the stomach.
RESULTS
Telomerase activity by TRAP was positive in all cases of adenocarcinoma, most cases of dysplasia, and many cases of normal mucosa. hTR in situ hybridization showed positive staining in the adenocarcinoma cells, dysplastic cells, a few cells in the proliferation zone of the normal mucosa, and a few infiltrated lymphocytes. hTERT showed positive staining in the same cells.
CONCLUSIONS
Telomerase is expressed in most cases of dysplastic lesions and is thought to be acquired in the early steps of carcinogenesis. The expression is noted in a few cells of the normal proliferative zones and the infiltrated lymphocytes, emphasizing the importance of in situ detection of telomerase at the cell level.

Detection of Human Papillomavirus 16/18, 31/33/35 in Verrucous Carcinoma of the Larynx by In Situ Hybridization with Human Papillomavirus DNA Probes.: Ki Kwon Kim, Sang Sook Lee, June Sik Park, Seung Won Jin, U Ik Sohn; Korean J Pathol. 1990;24(2):95-102.

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Abstract PDF: Verrucous carcinoma of the larynx is a distinct and uncommon variant of well-differentiated squamous cell-carcinoma. The authors hybridized in situ a case of laryngeal verrucous carcinoma with DNA prebes to humman papillomavirus (HPV) 6, 11, 16, 18, 31, 33 and 35. The DNAs from the verrucous carcinoma of larynx hybridized with mixed DNA probes HPV-16/18 and 31/33/35. In addition, there was an evidence of HPV infection based on hybridization with mixed DNA probe HPV-6/11 in the adjacent papilloma tissue. By in situ DNA hybridization techniques, we clearly demonstrated human papillomavirus (HPV-16/18, and 31/33/35) related sequences in this neoplasm. These findings suggest the role of HPV-6/11 in the development of laryngeal papilloma ad HPV-16/18 and 31/33/35 probably on the progression to verrucous carcinoma.

Case Reports

Rasmussen's Encephalitis.: Na Rae Kim, Han Jae Joon, Yeon Lim Suh, Moon Hyang Lee; Korean J Pathol. 2001;35(5):455-460.

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Abstract PDF: We herein report a case of intractable epilepsy that occurred in a 7-year-old girl, which is consistent with radiological and clinicopathological hallmarks of Rasmussen's encephalitis. The patient showed characteristic primary unilateral involvement with secondary bilateral propagation. Microscopically, the cortical atrophy due to neuronal loss, intense GFAP-immunoreactive astrogliosis, neuronophagia, perivascular lymphocytic infiltration and microglial nodules was seen throughout the cortex and white matter. No viral inclusions were noted; no cytomegalovirus, herpes simplex virus or Epstein-Barr virus was found by in situ hybridization. Granular immunofluorescence for C4, C1q and IgG within the blood vessel walls was noted, and ultrastructurally, only nonspecific vascular injury was found. Rasmussen's encephalitis is a diagnosis of exclusion; it can be diagnosed by the combination of clinical manifestation, neuroimaging and characteristic pathologic features.

Interfollicular Hodgkin's Lymphoma: A case report.: Lee So Maeng, Kyung Mee Kim, You Mee Kang, Chang Seok Kang, Sang In Shim; Korean J Pathol. 1995;29(6):804-806.

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Abstract PDF: An unusual pattern of focal involvement of lymph nodes by Hodgkin's disease is described using the name Interfollicular Hodgkin's Disease. It is characterized by florid reactive follicular hyperplasia which overshadows involvement of the interfollicular zones by Hodgkin's desease. The importance of interfollicular Hodgkin's disease rests on its misdiagnosis as a benign lesion. We report a case of interfollicular Hodgkin's disease in a 34-year-old female patient in the inguinal area. The lymph nodes showed reactive follicular hyperplasia. The focal interfollicular spaces were invaded by Hodgkin's disease. The Reed-Stemberg cells stained positively with CD15 and CD30 antibody in the immunohistochemical stain. An in situ hybridization study looking for EBV was negative in these R-S cells.

Original Articles

Detection of Numerical Chromosomal Aberration in Squamous Cell Carcinoma of the Lung by In Situ Hybridization Using #17 Centromeric Probes.: Sang Sook Lee, Seong Beom Han, Soong Kook Park; Korean J Pathol. 1993;27(5):443-458.

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Abstract PDF: This study was carried out to understand the relationship between specific chromosome changes and their phenotypic consequences at the tissue level of human lung cancers. Then paraffin-embedded human lung squamous cell carcinoma samples were investigated for in evidence of genetic alterations, using chromosome 7 and 17-specific repetitive alpha-satellite DNA probes. In situ hybridization procedure with chromosome-specific DNA probes was optimized for use on formalin-fixed paraffin-embedded lung tissue sections. The chromosome index ranged from 1.10 to 1.88(median, 1.49) for chromosome 7 and 1.20 to 1.98(median, 1.69) for chromosome 17. Normal lymphocytes and stromal cells showed one or two chromosome signals per cell in most cases. All tumors showed three or more chromosome signals per cell with range of 16.0% to 80.6% of cancer cells(median, 50.9%) for chromosome 7 and 32.7% to 84.7%(median, 69.9%) for chromosome 17. The chromosome index did not correlate with the DNA content in most cases. Chromosomes 7 and 17 were either overrepresented or underrepresented when they were compared with corresponding DNA index determined by FCM. An increase in copy number, particularly of chromosome 7 was associated with a less favorable phenotype, including high nuclear grade. In addition, chromosome alterations were differentially expressed in the different areas of the same tissue section, correlating with histologic heterogeneity. These results suggest that chromosome polysomy can be reliably detected in tissue sections using in situ hybridization. There is a strong correlation between genotypic abnormalities and tumor phenotype in human lung cancer. This capability will prove to be an important tool for determining the underlying genetic basis for tumor development, tissue phenotype heterogeneity and progression by allowing genetic determination to be made on paraffin-embedded tissue sections where tumor histologic architecture is preserved.

Detection of Chromosome Aberration in Interphase Nuclei of Tumor Cells by Nonradioactive In Situ Hybridization Using Chromosome-specific Probes.: Sang Sook Lee; Korean J Pathol. 1993;27(6):573-580.

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Abstract PDF: An improved protocol for in situ hybridization(ISH) to routinely processed, paraffin-embedded tissue sections from lung carcinoma is presented. For this study, DNA probes for alpha-satellite chromosome 7 and 17 were used. The protocol to detect numerical chromosome aberrations involved treatment of sections with 1 M sodium thiocyanate prior to pepsin digestion, resulting in reproducible ISH reactions. The effect of avidin-biotin detection system. Four layer avidin methods and triple biotin methods, using avidin-PO, goat antiavidin, biotinylated antigoat IgG, avidin-PO or anti-biotin, biotinylated antirabbit IgG, avidin-PO, markedly enhanced the intensity of positive signals. More than 80% of the tumor and stromal cells showed distinct chromosome hybridization signals in 6 micrometer-thick sections. Lung carcinoma cells showed multiple chromosome signals(2~5 spots), contrasted by one or two signals in the stromal cells in the same section. These results suggest that chromosome polysomy can be reliably detected in tissue sections using in situ hybridization. This capability will prove to be an important tool for determining the underlying genetic basis for tumor development, tissue phenotype heterogeneity and progression by allowing genetic determination to be made on paraffin-embedded tissue sections where tumor histologic architecture is preserved.

Type IV Collagen mRNA Expression in Human Membranous Nephropathy.: Tae Sook Kim, Jung Yeon Kim, Hye Kyoung Hong, Hyun Soon Lee; Korean J Pathol. 1999;33(11):1047-1054.

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Abstract PDF: Human membranous nephropathy (MN) is morphologically characterized by subepithelial immune complex deposits and progressive thickening of glomerular basement membranes (GBM). Studies have suggested that the enhanced secretion of classical and novel type IV collagen chains in MN contributes to spike formation and the novel type IV collagen chain is particularly related to thickening of GBM. It is unclear whether the increased accumulation of extracellular matrix (ECM) proteins in GBM is due to the increased mRNA expression for type IV collagen in glomerular visceral epithelial cells (GECs). To answer this question, we analyzed seven renal biopsies of patients with idiopathic MN using in situ hybridization. In MN, the number of GECs expressing mRNA for alpha1(IV) collagen was 2.82+/-1.80/glomerular cross section (gcs), and the number expressing mRNA for alpha4(IV) collagen was 8.42+/-2.85/gcs. The number of GECs expressing mRNA for alpha4(IV) collagen was significantly larger than that of alpha1(IV) collagen mRNA. The expression of mRNA for these ECM proteins in normal controls was negligible. These results suggest that subepithelial immune complexes stimulate the gene expression of alpha1(IV) collagen and alpha4(IV) collagen in glomerular GECs which, in turn, increase the secretion of ECM proteins and contribute to the thickening of GBM in MN.

Correlation between Transforming Growth Factor-beta and Procollagen III with Regenerative Activity in Acute Liver Injury, and the Effect of Prostaglandin E2.: Nam Hoon Cho, Chan Il Park; Korean J Pathol. 1996;30(5):367-387.

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Abstract PDF: Transforming growth factor-beta (TGF-beta) plays an important role in hepatic fibrogenesis. It is thought to inhibit regeneration of the hepatocytes. The aim of this present study was to clarify the correlation of TGF-beta, collagen type III (PIIINP) and the regenerating activity of hepatocytes, and the effect of prostaglandin E2 (PgE2) on them in acute liver injury. Two hundred and sixteen male Sprague-Dawley rats, weighing 200g on average, were divided into six experimental groups and two control groups; group I-CCl4 only administration, group II-partial hepatectomy(PH) only, group III-PH following CCl4 administration, group IV-olive oil only administration, group V-sham operation, group VI-CCl4 administration with pretreatment of PgE2, group VII- PH with pretreatment of PgE2, and group VIII- PH following CCl4 administration with pretreatment of PgE2. Five rats were sacrificed at 12, 24, 36, 48, 96 and 168 hours after the administration of CCl4 or PH in each experimental group. The liver was tested with immunohistochemical stain for proliferating cell nuclear antigen (PCNA) and in situ hybridization for TGF-beta. Radioimmunoassay for serum PIIINP was also performed. The results were as follows: TGF-beta was expressed mainly in the perisinusoidal cells and periportal mesenchymal cells. The TGF-beta positive cells were most numerous in the combined group of CCl4 plus PH. TGF-beta expression tended to have an inverse relation, with the PCNA index in all experimental groups. The PCNA index was highest in the CCl4 only group and lowest in the combined group of CCl4 plus PH. The PH only group showed a peak PCNA index at 48 hours. In the CCl4 only group and the combined group of CCl4 plus PH, serum PIIINP appeared to increase at 12 hours or more after the expression of hepatic TGF-beta. Pretreatment of PgE2 revealed that TGF-beta precipitously disappeared at 48-96 hours after insult. PgE2 influenced the vanishing period, not the emerging time of TGF-beta and had a remarkable effect on the amount of TGF-beta especially in the PH following CCl4 administration group, which resulted in significant accentuation of PCNA indices. In conclusion, PH of the prior injured liver induces a marked increase of TGF-beta followed by a significant suppression of regeneration of the remaining liver, and PgE2 overtly suppresses the expression of TGF-beta.

Comparing Fluorescence In Situ Hybridization and Immunohistochemistry to Determine the HER-2/neu Status in Breast Carcinoma.: Kyeongmee Park, Jungyoen Kim, Sungjig Lim; Korean J Pathol. 2002;36(4):243-248.

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Abstract PDF: BACKGROUND
Identification of HER-2/neu status is important in predicting the response to specific chemotherapy in breast carcinoma patients and HER-2/neu status is associated with poor clinical outcome even with systemic chemotherapy. Introduction of fluorescence in situ hybridization (FISH) allows an accurate assessment of the level of gene amplification with information about distribution of gene copies in histologic sections.
METHODS
HER-2/neu status was performed on paraffin sections of 176 primary breast carcinomas by FISH, using PathVysion and by immunohistochemistry (IHC), using HercepTest. The results of HER-2/neu amplification was compared with clinical and pathological prognostic factors.
RESULTS
HER-2/neu amplification and overexpression were detected in 51 tumors (29.0%) by FISH and 32 tumors (18.2%) by IHC. The results of each method agreed with each other in 157 tumors (concordance: 89.2%, kappa=0.783). HER-2/neu amplification was associated with poor nuclear grade, marked nuclear pleomorphism, and presence of the combined ductal carcinoma in situ in the invasive ductal carcinomas as well as Van Nuys grade of the ductal carcinoma in situ component (p<0.05).
CONCLUSIONS
The comparison of FISH and IHC demonstrated an excellent correlation of HER-2/neu overexpression 2+ and 3+ with gene amplification. However, FISH may be a more accurate and reliable method for negative and 1+ cases. HER-2/neu amplification proves to be of prognostic relevance.

Application of Epstein-Barr Virus Cell Lines (CCL85 EB-3) in Performing the EBER mRNA In Situ Hybridization as a Positive Control.: Sung Sook Kim, Woon Sup Han, Joo Young Suh, Joo Ryung Huh; J Pathol Transl Med. 1996;7(1):38-43.

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Abstract PDF: Epstein-Barr virus(EBV) is associated with a wide spectrum of benign and malignant disorders including leukoplakia, Hodgkin's lymphoma, central nervous system lymphoma, peripheral T cell lymphoma and nasopharyngeal undifferentiated carcinoma. There are several distinctive aspects of biology of the virus that are important in investigation of virus in clinical specimens. The abundant expression of the EBER mRNA transcripts makes possible the sensitive detection of latent expression in EBV-associated tumors. Although there has been a dramatic increased interest in the direct characterization of EBV in clinical specimens, there have been few studies about the effective and reliable positive controls in performing in situ hybridization technique for EBV, especially on paraffin-embedded tissue. We applied Burkitts lymphoma cell line as positive control in EBV in hydridization using Oncor Kit. The cell block of Burkitt lymphoma cell line(CCL85 EB-3) showed strong and specific positivity for EBER in situ in nuclei of EBV infected cells.

HER-2/neu Oncogene Amplification by Chromogenic in situ Hybridization and Immunohistochemical Expression of Topoisomerase II-alpha in the Breast Cancer.: Tae Jin Lee, Hyung Goon Oh, Gui Young Kwon, Mi Kyung Kim, Eon Sub Park, Jae Hyung Yoo; Korean J Pathol. 2003;37(1):26-34.

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Abstract PDF: BACKGROUND
Amplifications of the HER-2/neu oncogene and the Topoisomerase II-alpha gene are important determiners of the response to chemotherapy in the breast cancer. For detecting HER-2/neu amplification, fluorescent in situ hybridization and immunohistochemistry are currently regarded as standard methods. Chromogenic in situ hybridization (CISH) is investigated as a new modification of in situ hybridization. The purpose of this study is to compare the efficacy of CISH and immunohistochemistry (IHC) in detecting HER-2/neu oncogene amplification and to investigate the prognostic significance of the HER-2/neu oncogene and the Topoisomerase II-alpha gene in breast cancer.
METHODS
Using CISH and IHC the amplifications and protein expressions of the HER-2/neu oncogene were studied on paraffin sections of 43 infiltrating duct carcinomas. The expression of the Topoisomerase II-alpha gene was studied immunohistochemically.
RESULTS
Of the 43 infiltrating duct carcinomas, amplifications of the HER-2/neu oncogene by CISH were observed in 8 cases (18.6%), and the HER-2/neu protein was deemed overexpressed by IHC in 9 cases (20.9%). The amplifications of the HER-2/neu oncogene showed a statistically significant correlation with tumor size, histological grade, and the Topoisomerase II-alpha index. The Topoisomerase II-alpha index showed a statistically significant correlation with tumor size, lymph node status, stage, histologic grade, and estrogen receptor status.
CONCLUSIONS
CISH is a useful alternative for determining HER-2/neu amplification, especially for confirming the immunohistochemical staining results. HER-2/neu amplification and the Topoisomerase II-alpha gene index may be prognostic factors of breast cancer.

Ultrastructural In Situ Hybridization Method for Detection of Oncogenic Viruses.: Keun Hong Kee, Chul Hee Choi, Ho Jong Jeon; Korean J Pathol. 1996;30(8):687-698.

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Abstract PDF: In situ hybridization (ISH) is a standard method for localizing DNA or RNA sequences in tissue or cell preperation. The technique was developed at the electron-microscopic level, and enables the precise subcellular localization. A method was developed for detection of specific viral DNA. We have tested various methods and technique to detect specific viral DNA through ISH at the electron microscopic level. Postembedding method of ultrastructural ISH was developed and successfully applied for the detection of human papillomavirus type 16 in squamous cell carcinoma of the uterine cervix and Epstein-Barr virus in EBV-infected leukemia cell line. The following results are made. The best results were obtained using 0.2% glutaraldehyde and 4% paraformaldehyde fixed tissue or cell block. The labelling was best observed on Unicryl resin and Lowicryl K4M resin sections. Epon sections showed no reactivity. Thin sections of Unicryl resin were more easier than Lowicryl K4M resin. Enzymatic predigestion with proteinase K, pepsin and trypsind gave good results. However, high concentration of these produce poor results due to excessive destruction of the cellular components. Alkali treatment with 0.5N sodium hydroxide produced successful results in denaturation of target DNA. The labelling density of gold particles was independent of incubation time or temperature in hybridization step. The viral DNA labelling was localized mainly within the nucleus, both within and at the edge of electron dense regions, and below the nuclear membrane. And the labelling was seen in the form of a dense, roughly spherical shape. In conclusion, the best results are obtained by the conditions that tissue fixed by 0.2% glutaraldehyde and 4% paraformaldehyde solution, embedded with Unicryl resin, protein denaturation by 0.1ul/ml proteinase K, DNA denaturation by 0.5N sodium hydroxide, and reaction with DNA probe.

Epstein-Barr Viral RNA(EBERs) Expression in Conventional Malignant Lymphoma and Polymorphic Reticulosis of Upper Aerodigestive Tract.: Do Youn Park, Kang Suek Suh, Sun Kyung Lee; Korean J Pathol. 1996;30(10):893-902.

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Abstract PDF: The author examined the immunophenotype and expression of Epstem-Barr virus RNA (EBERs) used in the situ hybridization technique in 20 cases of conventional malignant lymphoma and 28 cases of polymorphic reticulosis and malignant lymphoma with features of polymorphic reticulosis occured in the upper aerodigestive tract including the upper digestive tract(palatine tonsil), and upper respiratory tract(nasal cavity, paranasal sinus, nasopharynx). The results obtained were summarized as followings; 1. The favorable site of malignant lymphoma that occured in the upper aerodigestive tract was in the palatine tonsil(11 out of 20 cases, 55%), those of polymorphic reticulosis and malignant lymphoma with features of polymorphic reticulosis were nasal cavity and nasopharynx(19 out of 28 cases, 78%). 2. The immunophentype of malignant lymphoma that occured in the upper aerodigestive tract was mostly B-cell phenotype (15 out of 20 cases, 75%), and that of polymorphic reticulosis and malignant lymphoma with features of polymorphic reticulosis was predominantly T-cell phenotype(22 out of 28 cases, 79%). 3. The EBERs positivity of malignant lymphoma that occured in the upper aerodigestive tract was 25%(5 out of 20 cases), but that of polymorphic reticulosis and malignant lymphoma with features of polymorphic reticulosis was 57%(16 out of 28 cases). 4. The positive cases for EBERs revealed angiocentricity with necrosis(16 out of 21 cases, 76%), predominantly T-cell phenotype(19 out of 21 cases, 90%), and favorably involved the nasal cavity and nasopharynx(16 out of 21 cases, 76%). Based on the above results, it was concluded that polymorphic reticulosis and malignant lymphoma with features of polymorphic reticulosis that occurred in the upper respiratory tract was an EBV-positive angiocentric T-cell lymphoma favorably involving the nasal cavity and nasopharynx.

Liver Cell Dysplasia: Analysis of 141 cases with reference to histopathologic Characterization and proliferative activity.: Sang Yong Song, Yong Il Kim; Korean J Pathol. 1992;26(4):338-347.

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Abstract PDF: Liver cell dysplasia of Anthony(LCD) is a common association in hepatocellular carcinoma(HCC)-bearing liver and has been regarded as a premalignant condition with strong linkage to hepatitis B virus infection and cirrhosis. A total of 189 surgically resected livers [HCC(168 cases), cholangiocarcinoma(3 cases), metastatic carcinoma(3 cases), and non-neoplastic lesions(15 cases)] were reviewed to elucidate the nature of LCD by means of light microscopic examination, in situ hybridization method for HBV DNA and expression of proliferatin cell nuclear antigen(PCNA) using immunohistochemical technique. LCD was present in 141 cases(74.6%), and its prevalence rate was independently significant in HCCs with or without cirrhosis than other groups. There was no difference in mean age, although LCD-positive group was younger than its negative counterpart. Association rate of LCD in HCC-cirrhosis group was statistically significant than the non-cirrhotic group, and higher histological grading of LCD was correlated well with wider distribution pattern and clustering. Seropositivity of HBsAg was not correlated with presence of LCD or with histological grading. In situ hybridization techique using HBV DNA probe demonstrated fine granular stainable particles even in LCD cells. Immunohistochemical study for PCNA revealed that the proliferative activity of LCD was lower than that of the cirrhotic cell. With the above results it is concluded that LCD reflects neither a regenerating condition nor a premalignant lesion but suggest a reactive change.

Case Reports

Complete Hydatidiform Mole with a Coexisting Fetus: A case report .: Yi Kyeong Chun, Hye Sun Kim, Yee Jeong Kim, Hy Sook Kim, Soo Kyung Choi, So Yeon Park, Sung Ran Hong; Korean J Pathol. 2000;34(9):673-676.

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Abstract PDF: Twin conceptus of a complete hydatidiform mole (CHM) and a normal fetus are rare but are important because of diagnostic difficulty, problems related to twin pregnancy, and high risk of persistent gestational trophoblastic tumor. Recently, we experienced one case of twin pregnancy consisting of a CHM and a normal fetus. A 26-year-old woman complained of vaginal bleeding. She had evidences of pregnancy-induced hypertension. A male fetus was delivered at 20 gestational weeks. The placenta demonstrated vesicles of molar change separated from normal placenta. Microscopically, the molar villi disclosed diffuse hydropic swelling with circumferential trophoblastic proliferation. DNA flow cytometric analysis showed diploid patterns in both molar and normal placental tissues. Fluorescent in situ hybridization in paraffin-embedded tissue presented that normal placental villi hybridized with X- and Y-chromosome probes (46, XY), while molar villi hybridized with X-chromosome only (46, XX). Thus, dizygotic twinning was confirmed because sex differences were shown between molar villi and normal placental villi. Follow up beta-hCG was within normal range after delivery.

Intraneural Perineurioma in the Tongue: A Case Report.: Jun Kang, Shin Kwang Khang, Jene Choi, Jeong Won Kim, Eul Ju Seo, Bu kyu Lee, Eunsil Yu; Korean J Pathol. 2007;41(1):51-54.

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Abstract PDF: We report a case of an intraneural perineurioma that developed in an unusual location, the tongue. A 16-year-old male presented with a 1 cm sized protruding submucosal mass in his tongue without any sensory or motor signs or symptoms. The mass was excised. The mucosa was intact, with an ill-defined firm mass measuring 1.0 x 0.8 x 0.6 cm in the submucosa and muscle. The cut surface of the mass was pinkish gray and fibrotic. Microscopically, the mass contained tortuous and thickened peripheral nerve bundles in the submucosa, showing onion bulb like structures. The onion bulb like structures consisted of centrally located S-100 protein positive Schwann cells surrounded by Glut-1 positive perineurial cells. The FISH study did not reveal any genetic aberrations in chromosome 22.

Original Articles

Expression of MMP-2, MT1-MMP, and TIMP-2 mRNA in Breast Carcinomas.: Dong Won Kim, So Young Jin, Dong Wha Lee; Korean J Pathol. 2003;37(6):400-406.

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Abstract PDF: BACKGROUND
The activation of proMMP-2 is induced by membrane type 1-matrix metalloproteinase (MT1-MMP), but inhibited by tissue inhibitors of matrix metalloproteinase type 2 (TIMP-2). This study was carried out to establish the pattern of mRNA expression of MMP-2, MT1-MMP, and TIMP-2 in breast carcinomas.
METHODS
Seventy-nine cases of invasive ductal carcinoma, 10 of ductal carcinoma in situ, and 10 of fibrocystic disease as a control were analysed for the expression of MMP-2, MT1-MMP, and TIMP-2 mRNA, using in situ hybridization. Correlations of the results with the clinical stage, tumor size, nodal status, and nuclear grade were analysed.
RESULTS
The expression rates of MMP-2, MT1-MMP, and TIMP-2 mRNA in invasive ductal carcinoma were 68%, 73%, and 56%, respectively. They were localized to both stromal and tumor cells, but mainly in the latter. The MMP-2 mRNA expression was significantly correlated with the clinical stage (p < 0.05), while the expression of TIMP-2 mRNA was inversely correlated with clinical stage and tumor size(p < 0.05). Significant positive correlations between MMP-2 and MT1-MMP expressions, along with inverse relationships between MMP-2 and TIMP-2, and between TIMP-2 and MT1-MMP, were also found. CONCLUSIONS: MMP-2 and TIMP-2 mRNA expressions might be useful as one of a range of prognostic parameters in breast carcinoma patients.

Molecular Cloning of Novel Genes Related to the Craniofacial Development of Human Embryo.: Young Jun Lee, Tak Soo Go, Hyung Wook Han, Sang Shin Lee, Eun Cheol Kim, Yeon Sook Kim, Suk Keun Lee, Je G Chi; Korean J Pathol. 2000;34(12):961-971.

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Abstract PDF: In order to obtain novel genes for craniofacial development of human, molecular cloning and sequencing were performed and followed by in situ hybridization in tissue sections. Subtracted cDNA library of craniofacial tissue from 8 weeks old human embryo was made by the subtraction with cDNA of RHEK cells. A total of 231 clones were obtained and their partial sequence data disclosed that 214 clones were nonredundant in Genebank search. We have done in situ hybridization screening on the craniofacial sections of a 10 weeks old human fetus, and found significant positive reaction in 30 clones. Depending on the cell type of similar developmental origin, the positive reactions could be divided into four groups: first group showed an intense positive reaction in neural tube, ganglion, and a part of peripheral nerve tissue, second group relatively diffuse positive reaction in neural tube, cartilage, epithelium, and muscle, third group localized positive reaction in nerve, and muscle, and fourth group positive reaction in almost all kinds of cells of craniofacial tissues. Although every clone showed different expression patterns in the craniofacial development, some of them showed intense mRNA expressions in the characteristic cell type. Because this study also aimed to test a screening methods to find out novel genes related to craniofacial development by the subtracted cDNA library and in situ hybridization, the intense positive reaction of a certain clone by in situ hybridization may indicate its role in the developmental processes. We presumed that 30 clones selected in this study are possibly important new genes for the development of human craniofacial structure.

Elafin Expression in Oral Lichen Planus.: Sang Shin Lee, Suk Keun Lee; Korean J Pathol. 2004;38(1):15-22.

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Abstract PDF: BACKGROUND
Elafin is a potent anti-elastase in human saliva, and is supposed to play a role in preventing oral ulceration. The expression of elafin was observed in the oral lichen planus (OLP), one of the most common noninfectious oral mucosal diseases, which frequently manifests as extensive ulceration on the involved oral mucosa.
METHODS
50 OLP, 10 oral leuko-plakia, 3 inflammatory oral ulcers, and 3 normal oral mucosa cases were fixed with 10% buffered formalin, and immunohistochemically stained with monoclonal elafin antibody. Representative specimens were fixed with 4% paraformaldehyde, and RNA in situ hybridization, with an elafin RNA probe, was performed.
RESULTS
With both the immunohistochemistry and RNA in situ hybridization the expression of elafin was more decreased in the OLP compared to the normal mucosa, while in the hyperplastic epithelium of the leukoplakia and inflammatory ulcers the expressions of elafin was more intense. In the thin epithelia of the reticular and atrophic OLPs the expressions of elafin were reduced compared to the normal mucosa, and became almost negative in the epithelium of the erosive OLP.
CONCLUSIONS
These data suggested that the extensive ulceration of the OLP was closely relevant to the reduced expression of elafin in the involved epithelium

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