Journal of Pathology and Translational Medicine

Original Articles

Diagnostic Utility of a Clonality Test for Lymphoproliferative Diseases in Koreans Using the BIOMED-2 PCR Assay: Young Kim, Yoo Duk Choi, Chan Choi, Jong-Hee Nam; Korean J Pathol. 2013;47(5):458-465. Published online October 25, 2013; DOI: https://doi.org/10.4132/KoreanJPathol.2013.47.5.458

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Background

A clonality test for immunoglobulin (IG) and T cell receptor (TCR) is a useful adjunctive method for the diagnosis of lymphoproliferative diseases (LPDs). Recently, the BIOMED-2 multiplex polymerase chain reaction (PCR) assay has been established as a standard method for assessing the clonality of LPDs. We tested clonality in LPDs in Koreans using the BIOMED-2 multiplex PCR and compared the results with those obtained in European, Taiwanese, and Thai participants. We also evaluated the usefulness of the test as an ancillary method for diagnosing LPDs.

Methods

Two hundred and nineteen specimens embedded in paraffin, including 78 B cell lymphomas, 80 T cell lymphomas and 61 cases of reactive lymphadenitis, were used for the clonality test.

Results

Mature B cell malignancies showed 95.7% clonality for IG, 2.9% co-existing clonality, and 4.3% polyclonality. Mature T cell malignancies exhibited 83.8% clonality for TCR, 8.1% co-existing clonality, and 16.2% polyclonality. Reactive lymphadenitis showed 93.4% polyclonality for IG and TCR. The majority of our results were similar to those obtained in Europeans. However, the clonality for IGK of B cell malignancies and TCRG of T cell malignancies was lower in Koreans than Europeans.

Conclusions

The BIOMED-2 multiplex PCR assay was a useful adjunctive method for diagnosing LPDs.

Citations

Citations to this article as recorded by

Infectious mononucleosis complicated by transitory Epstein-Barr virus infection of T and natural killer cells
Yanlin Zhang, JianLan Xie, Yuanyuan Zheng, XiaoGe Zhou
Journal of Hematopathology.2024; 17(3): 129. CrossRef
Experiencia en el uso de protocolos Biomed-2 para el estudio de reordenamientos de TCR e inmunoglobulinas en proliferaciones linfoides en el Instituto Nacional de Cancerología, Colombia
Nicolás Villamizar-Rivera, Natalia Olaya
Biomédica.2022; 42(Sp. 1): 64. CrossRef
Enhancing diagnosis of T-cell lymphoma using non-recombined T-cell receptor sequences
Yi-Lin Chen, Chung-Liang Ho, Chen-Yan Hung, Wan-Li Chen, Chen Chang, Yi-Hsin Hou, Jian-Rong Chen, Pin-Jun Chen, Nan-Haw Chow, Wenya Huang, Ya-Ting Hsu, Tsai-Yun Chen, Tsunglin Liu
Frontiers in Oncology.2022;[Epub] CrossRef
The utility and limitations of B- and T-cell gene rearrangement studies in evaluating lymphoproliferative disorders
Hadrian Mendoza, Christopher A. Tormey, Henry M. Rinder, John G. Howe, Alexa J. Siddon
Pathology.2021; 53(2): 157. CrossRef
Combined detection of lymphocyte clonality and MALT1 translocations in bronchoalveolar lavage fluid for diagnosing pulmonary lymphomas
Takashi Kido, Hiroshi Ishimoto, Hiroshi Ishii, Kanako Hara, Mutsumi Ozasa, Hiroki Kawabata, Toshinori Kawanami, Yu Suzuki, Hiroki Yoshikawa, Atsuko Hara, Noriho Sakamoto, Nobuhiro Matsumoto, Chiharu Yoshii, Junya Fukuoka, Masaki Fujita, Masamitsu Nakazato
Scientific Reports.2021;[Epub] CrossRef
Differentiation of lymphocytic‐plasmacytic enteropathy and small cell lymphoma in cats using histology‐guided mass spectrometry
Sina Marsilio, Shelley J. Newman, James Scot Estep, Paula R. Giaretta, Jonathan A. Lidbury, Emma Warry, Andi Flory, Paul S. Morley, Katy Smoot, Erin H. Seeley, Matthew J. Powell, Jan S. Suchodolski, Jörg M. Steiner
Journal of Veterinary Internal Medicine.2020; 34(2): 669. CrossRef
T-Cell Receptor Rearrangements Determined Using Fragment Analysis in Patients With T-Acute Lymphoblastic Leukemia
Hyerim Kim, In-Suk Kim, Chulhun L. Chang, Sun-Young Kong, Young Tak Lim, Seom Gim Kong, Eun Hae Cho, Eun-Yup Lee, Ho-Jin Shin, Hyeon Jin Park, Hyeon-Seok Eom, Hyewon Lee
Annals of Laboratory Medicine.2019; 39(2): 125. CrossRef
Monitoring immunoglobulin heavy chain and T‑cell receptor gene rearrangement in cfDNA as minimal residual disease detection for patients with acute myeloid leukemia
Ling Zhong, Jiao Chen, Xiaobing Huang, Yanxing Li, Tao Jiang
Oncology Letters.2018;[Epub] CrossRef
Molecular pathology diagnosis of diffuse large B cell lymphoma using BIOMED-2 clonal gene rearrangements
Saeid Ghorbian
Annals of Diagnostic Pathology.2017; 29: 28. CrossRef
Improved clonality detection in B‐cell lymphoma using a semi‐nested modification of the BIOMED‐2 PCR assay for IGH rearrangement: A paraffin‐embedded tissue study
Yuma Sakamoto, Ayako Masaki, Satsuki Aoyama, Shusen Han, Kosuke Saida, Kana Fujii, Hisashi Takino, Takayuki Murase, Shinsuke Iida, Hiroshi Inagaki
Pathology International.2017; 67(9): 453. CrossRef
The prognostic significance of monoclonal immunoglobulin gene rearrangement in conjunction with histologic B‐cell aggregates in the bone marrow of patients with diffuse large B‐cell lymphoma
Yoon Ah Cho, Woo Ick Yang, Jae‐Woo Song, Yoo Hong Min, Sun Och Yoon
Cancer Medicine.2016; 5(6): 1066. CrossRef
Nasal-type NK/T-cell lymphomas are more frequently T rather than NK lineage based on T-cell receptor gene, RNA, and protein studies: lineage does not predict clinical behavior
Mineui Hong, Taehee Lee, So Young Kang, Suk-Jin Kim, Wonseog Kim, Young-Hyeh Ko
Modern Pathology.2016; 29(5): 430. CrossRef
Long-term Tumor-free Survival With Untreated Primary Diffuse Large B-cell Lymphoma of the Tonsil
Xiaojing Zhang, Yuanyuan Zheng, Jianlan Xie, Jun Zhu, Yuqin Song, Xiaojing Teng, Wei Liu, Yi Ding, Yuhua Huang, Xiaoge Zhou
American Journal of Surgical Pathology.2015; 39(11): 1493. CrossRef
Evaluation diagnostic usefulness of immunoglobulin light chains (Igκ, Igλ) and incomplete IGH D-J clonal gene rearrangements in patients with B-cell non-Hodgkin lymphomas using BIOMED-2 protocol
S. Ghorbian, I. Jahanzad, G. R. Javadi, E. Sakhinia
Clinical and Translational Oncology.2014; 16(11): 1006. CrossRef

Comparison of Three BRAF Mutation Tests in Formalin-Fixed Paraffin Embedded Clinical Samples: Soomin Ahn, Jeeyun Lee, Ji-Youn Sung, So Young Kang, Sang Yun Ha, Kee-Taek Jang, Yoon-La Choi, Jung-Sun Kim, Young Lyun Oh, Kyoung-Mee Kim; Korean J Pathol. 2013;47(4):348-354. Published online August 26, 2013; DOI: https://doi.org/10.4132/KoreanJPathol.2013.47.4.348

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Abstract PDF

Background

Recently, BRAF inhibitors showed dramatic treatment outcomes in BRAF V600 mutant melanoma. Therefore, the accuracy of BRAF mutation test is critical.

Methods

BRAF mutations were tested by dual-priming oligonucleotide-polymerase chain reaction (DPO-PCR), direct sequencing and subsequently retested with a real-time PCR assay, cobas 4800 V600 mutation test. In total, 64 tumors including 34 malignant melanomas and 16 papillary thyroid carcinomas were analyzed. DNA was extracted from formalin-fixed paraffin embedded tissue samples and the results of cobas test were directly compared with those of DPO-PCR and direct sequencing.

Results

BRAF mutations were found in 23 of 64 (35.9%) tumors. There was 9.4% discordance among 3 methods. Out of 6 discordant cases, 4 cases were melanomas; 3 cases were BRAF V600E detected only by cobas test, but were not detected by DPO-PCR and direct sequencing. One melanoma patient with BRAF mutation detected only by cobas test has been on vemurafenib treatment for 6 months and showed a dramatic response to vemurafenib. DPO-PCR failed to detect V600K mutation in one case identified by both direct sequencing and cobas test.

Conclusions

In direct comparison of the currently available DPO-PCR, direct sequencing and real-time cobas test for BRAF mutation, real-time PCR assay is the most sensitive method.

Citations

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Preoperative BRAFV600E mutation detection in thyroid carcinoma by immunocytochemistry
Kristine Zøylner Swan, Stine Horskær Madsen, Steen Joop Bonnema, Viveque Egsgaard Nielsen, Marie Louise Jespersen
APMIS.2022; 130(11): 627. CrossRef
Strategy to reduce unnecessary surgeries in thyroid nodules with cytology of Bethesda category III (AUS/FLUS): a retrospective analysis of 667 patients diagnosed by surgery
Yong Joon Suh, Yeon Ju Choi
Endocrine.2020; 69(3): 578. CrossRef
A new primer construction technique that effectively increases amplification of rare mutant templates in samples
Jr-Kai Huang, Ling Fan, Tao-Yeuan Wang, Pao-Shu Wu
BMC Biotechnology.2019;[Epub] CrossRef
BRAF and NRAS mutations and antitumor immunity in Korean malignant melanomas and their prognostic relevance: Gene set enrichment analysis and CIBERSORT analysis
Kyueng-Whan Min, Ji-Young Choe, Mi Jung Kwon, Hye Kyung Lee, Ho Suk Kang, Eun Sook Nam, Seong Jin Cho, Hye-Rim Park, Soo Kee Min, Jinwon Seo, Yun Joong Kim, Nan Young Kim, Ho Young Kim
Pathology - Research and Practice.2019; 215(12): 152671. CrossRef
The association between dermoscopic features and BRAF mutational status in cutaneous melanoma: Significance of the blue-white veil
Miquel Armengot-Carbó, Eduardo Nagore, Zaida García-Casado, Rafael Botella-Estrada
Journal of the American Academy of Dermatology.2018; 78(5): 920. CrossRef
Comparison of Five Different Assays for the Detection of BRAF Mutations in Formalin-Fixed Paraffin Embedded Tissues of Patients with Metastatic Melanoma
Claire Franczak, Julia Salleron, Cindy Dubois, Pierre Filhine-Trésarrieu, Agnès Leroux, Jean-Louis Merlin, Alexandre Harlé
Molecular Diagnosis & Therapy.2017; 21(2): 209. CrossRef
Validation of an NGS mutation detection panel for melanoma
Anne Reiman, Hugh Kikuchi, Daniela Scocchia, Peter Smith, Yee Wah Tsang, David Snead, Ian A Cree
BMC Cancer.2017;[Epub] CrossRef
Transformation to Small Cell Lung Cancer of Pulmonary Adenocarcinoma: Clinicopathologic Analysis of Six Cases
Soomin Ahn, Soo Hyun Hwang, Joungho Han, Yoon-La Choi, Se-Hoon Lee, Jin Seok Ahn, Keunchil Park, Myung-Ju Ahn, Woong-Yang Park
Journal of Pathology and Translational Medicine.2016; 50(4): 258. CrossRef
Immunohistochemistry with the anti-BRAF V600E (VE1) antibody: impact of pre-analytical conditions and concordance with DNA sequencing in colorectal and papillary thyroid carcinoma
Katerina Dvorak, Birte Aggeler, John Palting, Penny McKelvie, Andrew Ruszkiewicz, Paul Waring
Pathology.2014; 46(6): 509. CrossRef

Comparison of Detecting Methods of BK Virus Infection in Patients with Renal Allograft Recipients.: Sung Hak Lee, Youn Jun Park, Chul Woo Yang, Yong Soo Kim, In Sung Moon, Chang Suk Kang, Yeong Jin Choi; Korean J Pathol. 2010;44(6):636-641.; DOI: https://doi.org/10.4132/KoreanJPathol.2010.44.6.636

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Abstract PDF

BACKGROUND
BK virus nephropathy (BKVN) is an emerging problem as a consequence of the use of potent immunosuppressive agents. Because optimal detection methods for the diagnosis of BKVN are required clinically, we compared the results of renal allograft biopsy, urine cytology, and urine and blood viral loads.
METHODS
Four hundred sixty two case notes from 2004 to 2009 at Seoul St. Mary's Hospital were reviewed. During that period, 286 cases of urine cytology for decoy cells, 938 cases of urine BKV reverse transcription-polymerase chain reaction (RT-PCR), and 1,029 cases of blood BKV RT-PCR were performed. All diagnostic methods were performed in 85 cases.
RESULTS
A histological diagnosis of BKVN was made in 2.4% of cases (11/462). Urine cytology for decoy cells was positive in 26.2% (75/286). BKV RT-PCR revealed viruria in positivity of 22.1% (207/938) and viremia in 5.2% (54/1,029). In cases of BKVN, the sensitivities of urine and blood BKV RT-PCR were all 100% and the specificities were 69% and 94.5%, respectively. In cases with positive urine decoy cells, the sensitivities of urine and blood BKV RT-PCR were 50% and 27.7%, with specificities of 77.7% and 100%, respectively.
CONCLUSIONS
BKV screening by RT-PCR assays may be a clinically useful noninvasive test to identify renal recipients with concurrent BKVN.

Citations

Citations to this article as recorded by

Prevalence of BK Virus among Iranian Renal Transplant Recipients: A Systematic Review and Meta-Analysis
Mohsen Ebrahimi, Alireza Mohebbi, Mohammad Mostakhdem Hashemi, Mobina Ashrafi Shahmirzadi
Journal of Clinical and Basic Research.2020; 4(4): 50. CrossRef
Asymptomatic hematuria associated with urinary polyomavirus infection in immunocompetent patients
Sung Hak Lee, Sung Hoo Hong, Ji Youl Lee, Tae Kon Hwang, Kyoung Suk Kim, Hyoungnam Lee, Yeong Jin Choi
Journal of Medical Virology.2014; 86(2): 347. CrossRef

Detection of HBV DNA in Needle Biopsied Paraffin Embedded Liver Tissues of Chronic Hepatitis B Patients by PCR: Comparison with Serological and Immunohistochemical Studies.: Hye Soo Lee, Kahng Yeul Oh, Joo Heon Kim, Yoon Jeong Kim, Sam Im Choi, Dong Geun Lee, Sang Ho Kim; Korean J Pathol. 1996;30(6):495-504.

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Abstract PDF: In this study, the prevalence of Hepatitis B virus(HBV) DNA in the needle biopsied paraffin embedded liver tissues of chronic hepatitis B patients by rapid nested PCR was examined. DNA was extracted by NaOH with boiling, and amplified by rapid air thermocycler with glass capillary tubes and nested PCR with two primer sets specific for the surface and the core genes of HBV. The PCR results were compared to that of serum HBeAg, serum HBV DNA by dot blot hybridization with a radioactive DNA probe, and tissue immunohistochemical (HBsAg/ HBcAg) studies. Among 44 patients with chronic hepatitis with serum HBsAg positivity, HBV DNA could be detected by PCR in 43 liver tissues (98%). This results were comparable to the positive rates of 94%(31/33) for serum HBV DNA, 80%(35/44) for serum HBeAg, and 59%(26/44) and 75%(33/44) for tissue HBsAg and HBcAg, respectively. The accordance rate between tissue PCR and serum DNA probe testing was 91%. The results indicate that HBV DNA detection by rapid nested PCR of paraffin embedded liver tissues by needle biopsy is a more sensitive method to detect the HBV DNA carrier than the serum HBeAg or tissue HBsAg/HBcAg status, and is well correlated with the result of serum HBV DNA probe testing. Therefore this method is a practical indicator for the diagnosis and replication status in retrospective analysis.

Polymerase Chain Reaction Analysis of Human Papillomavirus in Esophageal Squamous Cell Carcinoma with its Correlation to p53 mutation.: Wan Seop Kim, Eun Kyung Hong, In Kyu Kim, Moon Hyang Park, Jung Dal Lee; Korean J Pathol. 1996;30(11):1018-1026.

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Abstract PDF: HPV infection has been implicated strongly in the pathogenesis of human squamous cell carcinoma(SCC). We analysed a series of 28 surgically removed, invasive squamous cell carcinoma of the esophagus by polymerase chain reaction to detect HPV DNA using consensus primers and 8 type-specific primers of HPV (6, 11, 16, 18, 31, 33, 35, 51). HPV 6, 31, 35 or 51 DNA were detected in 20 out of 28 cases (71.4%) of the esophageal SCCs. HPV 51 was the most frequently detected type, occuring in 13 out of 28 cases (46.4%). p53 immunohistochemical staining was also performed to demonstrate any relationship to HPV DNA positivity. It showed positivity in 16 out of 28(57.1%) esophageal SCCs, and HPV DNA and p53 positivity were concurrently detected in 11 out of 28 cases of SCCs. There was no significant inverse relation between HPV DNA positivity and p53 expression(p>0.05). Our results supported HPV involvement in esophageal squamous cell carcinoma, and suggested there may be another pathway not related to the p53-binding pathway in the carcinogenesis of esophageal SCCs by HPV.

The p53 Mutation and DNA Ploidy in Human Metastatic Breast Cancer.: Seong Jin Cho, Ae Ree Kim, Nam Hee Won; Korean J Pathol. 1997;31(2):135-144.

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Abstract PDF: The p53 gene, one of the tumor suppressor genes, is believed to play an important role through mutation and overexpression in the progression of various human malignant tumors. To compare the p53 mutation status between the primary and metastatic lesions of breast cancers and to investigate the mutational pattern of p53, immunohistochemistry (IHC) and polymerase chain reaction and single strand conformational polymorphism (PCR-SSCP) were performed in 25 cases of breast cancers with paraffin embedded tissue. Mutant protein products or point mutation were detected through IHC or PCR-SSCP method. And flow cytometrical (FCM) analysis were performed in the same paraffin blocks to correlate the DNA ploidy and p53 mutation. The following results are summarized. 1. The detection of the p53 gene mutation and overexpression of the p53 protein were measured in 40% and 48%, respectively, in 25 primary tumors, either or both methods was detected in 64%. 2. A concordance rate of the p53 protein expression between the primary and metastatic lesions of 25 breast cancers was 100%, but the concordance rate of the p53 gene mutation was 72%. 3. The correlation between the p53 mutation and the DNA aneuploidy was not statistically significant (p=0.38) 4. A p53 mutation by IHC or PCR-SSCP was more frequently detected in grade III breast cancers than in grade I or II. 5. Among 5 to 9 exons of the p53 gene, exon 7 was the most frequent mutation spot in this study. 6. Additional mutation of the p53 gene was developed in the three metastatic lesions. With the above results it is suggested that the p53 protein overexpression by immunohistochemistry is not correlated with the p53 mutation by PCR-SSCP. The p53 mutation pattern between the primary and metastatic lesions are not idenitical and an additional point mutation can occur in the metastatic lesion. The DNA aneuploidy is more frequently detected in the cases with the p53 protein overexpression than in the p53 protein negative, but it is not statistically significant.

Prognostic Value of CD44v6 Isoform in Infiltrating Ductal Carcinoma of Breast.: Seung Cheol Lee, Yoon Kyung Sohn, Jung Sik Kwak, Woon Bok Jhung, Jung Wan Kim; Korean J Pathol. 1997;31(7):635-643.

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Abstract PDF: CD44 is a family of transmembrane glycoproteins involved in cell-cell and cell-matrix interactions. Expression of CD44 isofonns (splice variants) has been shown to be associated with poor prognosis in several human cancers. We evaluated the expression patterns of the CD44 isofortn (CD 44 splice variant v6) in infiltrating ductal carcinoma of the breast by immunohistochemical and RT-PCR method. Paraffin embedded blocks from seventy-five cases of mastectomized samples were analyzed immunohistochemically using monoclonoal antibody against CD44v6. CD44v6 was detected in fifty-seven cases (76%) of the tumor samples. Adjacent normal myoepithelial cells and ductal epithelial cells revealed focal positive reaction to CD44v6. Thirtytwo cases (80.0%) with lymph nodal metastasis revealed overexpression of CD44v6 monoclonal antibody, but twenty-five cases (71.4%) without nodal metastasis also showed positive reaction to CD44v6 monoclonal antibody, and there is no statistically significant value. Other prognostic factors of infiltrating ductal carcinoma, such as tumor size, histologic grade and hormonal receptors did not show any significant correlation with CD44v6 expression. The RT-PCR studies for 9 cases of infiltrating ductal carcinoma showed the same band patterns both in the normal and tumor tissues. From the above results, it is concluded that the expression of CD44v6 is not a valuable prognostic marker of infiltrating ductal carcinoma of breast.

Alteration of p53 Tumor Suppressor Gene in Hyperplastic Lesions and Adenocarcinomas of Uterine Endometrium - Immunohistochemistry and PCR-SSCP.: Eun Kyung Kim, Chan Kum Park, Gu Kong, Moon Hyang Park, Jung Dal Lee; Korean J Pathol. 1997;31(7):662-671.

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Abstract PDF: To investigate the role of the p53 gene in the development of endometrial adenocarcinoma and to study the relation between alteration of the p53 gene and histologic grade, the author studied the alteration of thep53 gene in hyperplastic lesions and adenocarcinomas of the uterine endometrium. The study was carried out with immunohistochemical stain and PCR-SSCP. The materials included ten cases of endometrial hyperplasia (five simple and five atypical complex) and 18 cases of endometrial adenocarcinoma. Overexpression of the p53 protein were found in one of five atypical complex hyperplasias (20%) and 11 of 18 adenocarcinomas (61.1%). The intensity of p53 overexpression appeared to have increasing tendency with higher histologic grade of adenocarcinomas. Among the II cases of adenocarcinoma that overexpressed p53 protien, five cases (45.5%) were found to have mutations by PCR-SSCP. One was grade 1 (20%), two were grade 11 (25%), and two were grade III (40%). The sites of mutation were three exon 8, one exon 5, and one exon 6. In conclusion, alteration of the p53 gene may paly a role in the development of endometrial adenocarcinoma and appears to occur as a late event in carcinogenesis.HHowever, inactivation of the p53 gene in early stage of tumor development cannot be excluded.

Characteristics of the Immortalized Human B-cells by Epstein-Barr Virus.: Ho Jong Jeon, Bong Nam Choi, Yoon Kyeong Oh; Korean J Pathol. 1997;31(9):832-846.

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Abstract PDF: Human lymphoblastoid B-cell lines immortalized by Epstein-Barr virus (EBV) were established from peripheral blood of patients with acute myeloblastic and chronic lymphocytic leukemia and chronic fatigue syndrome. The sera of patients with acute myeloblastic and chronic lymphocytic leukemia did not show antibodies to Epstein-Barr viral capsid antigen (VCA), but serum of a patient with chronic fatigue syndrome disclosed antibodies to VCA (IgG, IgM), and EBNA was demonstrated in peripheral blood mononuclear cells by polymerase chain reaction. The established cell lines were mature B-cell phenotypes with polyclonal proliferation in early passage and no evidence for commitment to other lineages. The immortalized cells by EBV were designated as CSUP-1 and CSUP-2 (from acute myeloblastic leukemia, FAB classification M2 and M1), CSUP-3 (from chronic lymphocytic leukemia) and CSUP-4 (from a patient with chronic fatigue syndrome). The CSUP-1, 2, 3, and 4 grew in suspension forming clumps with a doubling time of 38 to 49 hours. Colony formation was not recognized in plate. By light and electron microscopic examination, the immortalized cells showed features of lymphoblastoid to plasmacytoid lymphocytes, and multinucleated giant cells. The lymphoblastoid cells showed scanty cytoplasm with poorly developed organelles. Immunophenotypic analyses of CUSP-1, 2, 3, and 4 with monoclonal antibodies by flow cytometry showed B-cell phenotype with polyclonal proliferation in early passage. Epstein-Barr virus nuclear antigen was confirmed in the extracted DNAs from immortalized cells by polymerase chain reaction. DNA analysis showed a normodiploid stemline with a DNA index of 1.12. The established cells were strongly reactive for CD10, CD30 (Ki-1) in early passage, and bcl-2 and c-myc onco-protein in early and late passage. Karyotypic analysis of CSUP-1, 2, 3 and 4 showed 46, XY or 46, XX. No tumorigenesis in heterotransplanted SCID mouse was recognized. This immortalized cells by EBV should be a valuable cell lines to study the pathogenesis of EBV-related malignant lymphoma.

A Comparative Study of Immunohistochemistry and PCR-SSCP for Detection of p53 Mutation In Gastric Carcinoma.: Jong Soon Kim, Jae Hyuk Lee, Min Cheol Lee, Chang Soo Park, Sang Woo Juhung; Korean J Pathol. 1998;32(1):21-28.

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Abstract PDF: Mutation of the p53 tumor-suppressor gene in exons 4 through 9 was examined in 34 cases of primary advanced gastric cancer using PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) and the results were compared with p53 protein expression as determined by immunohistochemistry (IHC) using a monoclonal antibody(DO-1). p53 protein detected by IHC was observed in 14 cases (41.2%) and genotypic mutation detected by PCR-SSCP in exons 4-9 was observed in 13 cases (38.2%) One case showed an aberrant band on PCR-SSCP both in Exon 7 and Exon 8/9. p53 alteration detected by either IHC or PCR-SSCP was observed in 19 cases (55.9%), but only 8 cases (23.5%) showed both p53 mutation and protein expression. We also tried to obtain the correlation between relative intensity of the shifted bands on PCR-SSCP and percentage of positive cells by IHC, but a significant correlation was not seen between relative intensity of shifted bands on PCR-SSCP and positve cell ratio. A discrepancy between p53 protein expression and p53 mutation is observed in primary gastric carcinomas. The reason for this discrepancy are not apparent. However, examination of gastric carcinomas for mutations in other exons may identify a better correlation with protein overexpression. The results obtained in this study suggest that the negative reaction for p53 immunohistochemistry may not necessarily mean no genetic alteration of the p53 locus.

Histopathologic Analysis of Helicobacter pylori-associated Chronic Gastritis between cagA-positive and cagA-negative Strains.: Hun Kyung Lee, Gyeong Hoon Kang, Hwoon Yong Jung, On Ja Kim; Korean J Pathol. 1998;32(7):504-510.

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Abstract: Infection with Helicobacter pylori (H. pylori) leads to gastritis, but the majority of infected persons are asymptomatic, and it has been recently described that the ability of H. pylori to cause more severe disease is related to the presence of the cytotoxin-associated gene A (cagA). We investigated the prevalence of cagA-bearing strains in a group of H. pylori-positive gastritis, and compared the morphologic differences between cagA-positive and cagA-negative cases on H&E stained slides. Polymerase chain reaction (PCR) assays for detection of H. pylori and cagA of 62 gastric biopsy specimens were performed. All the slides were analyzed by the updated Sydney system. Forty eight (77.4%) were PCR positive for H. pylori and thirty four (54.8%) were positive for cagA. There were no significant differences in numbers of H. pylori, degree of infiltration of mononuclear cells and degree of atrophy between cagA-positive and cagA-negative groups. The rates of neutrophilic infiltration and intestinal metaplasia were significantly higher in cagA-positive group than in cagA-negative group. In conclusion, the detection of H. pylori by PCR method is more sensitive than that of microscopic examination and H. pylori strains possessing cagA are associated with an enhanced induction of severe gastritis.

Detection of Mycobacterium tuberculosis DNA in Ethanol-Fixed and Papanicolaou Stained Archival Materials.: Tae Sook Hwang, In Seo Park, Hye Seung Han, Jee Young Han, Young Bae Kim; Korean J Pathol. 1998;32(8):603-607.

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Abstract: Granuloma is a chronic inflammatory process associated with non-infectious agents or infectious diseases such as tuberculosis. It is well known that AFB staining, which has been used to determine the etiology of the granulomatous inflammation, lacks both sensitivity and specificity. Due to the slow growth rate of most pathogenic mycobacteria, culturing of organisms can take up to eight weeks. It is not uncommon for specific therapy to be delayed, or for an inappropriate treatment be given to patients without mycobacterial infections or with infections caused by atypical mycobacteria. Determination of the causative agent in Papanicolaou stained cytology specimens gives pathologists even more difficulties when only necrotic material has been aspirated from the center of the granuloma. In recent years, the use of a polymerase chain reaction for the amplification of DNA has appeared promising in terms of speed, efficiency, sensitivity, and specificity. Since a polymerase chain reaction permits the sensitive genetic analysis of small amounts of tissue, it is ideally suited to the genetic analysis of cytologic specimens. A polymerase chain reaction is easily performed on unfixed and unstained cells, however, an analysis of ethanol fixed and Papanicolaou-stained archival smears has also been described. We have recently established a method to detect Mycobacterium tuberculosis organism by a nested polymerase chain reaction with primers in the insertion sequence IS 6110, using cellular digests of ethanol-fixed and Papanicolaou-stained archival specimens aspirated from the lymph nodes, lungs, thyroid, etc. Inhibitors present in Papanicolaou stained material was removed by destaining the slides with 0.5% HCl solution for 10-30 minutes. Eight out of ten cases which have shown the epithelioid granulomas revealed a positive reaction and four out of ten cases which have shown lymphohistiocytic cells in a necrotic background without any evidence of granuloma revealed a positive reaction. This study showed that it was possible to employ a polymerase chain reaction to detect Mycobacterium tuberculosis in Papanicolaou stained archival cytology specimens.

Immunohistochemical Analysis of Transforming Growth Factor (TGF)-beta1 and TGF-beta Receptor II and Quantitative Analysis of TGF-beta1 mRNA during Multistep Hepatocarcinogenesis Induced by Diethylnitrosamine in Sprague-Dawley Rats.: Mee Yon Cho, Ju Han Lee, Yong Koo Kang, Nam Hee Won; Korean J Pathol. 1999;33(11):1009-1023.

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Abstract PDF: Transforming growth factor (TGF)-beta1 plays an important role in hepatocarcinogenesis and has been described as a useful tumor marker and one of the poor prognostic indicators in patients with hepatocellular carcinoma (HCC). To investigate the role and cellular localization of TGF-beta1 during multistep hepatocarcinogenesis we performed a quantitative analysis of TGF-beta1 mRNA and immunohistochemical expression of TGF-beta1 and TGF-beta receptor II (TGF-betarII) in female Sprague-Dawley rats. The experimental groups included neoplastic lesions produced by Solt-Farber's protocol, regenerating liver after partial hepatectomy, and normal control. Quantitative change of TGF-beta1 mRNA was analysed by competitive reverse-transcription polymerase chain reaction (RT-PCR). TGF-beta1 protein and TGF-betarII expression were evaluated by immunohistochemical stain. The discrete tumor nodules were detected on 14th day and then increased in number and size. Three HCCs were induced on 8th or 9th month. RT-PCR demonstrated TGF-beta1 mRNA band in all examples of the normal and regenerating liver, nodules and HCCs. Competitive RT-PCR displayed higher TGF-beta1 mRNA in nodules, HCCs and regenerating liver than in normal controls. Hepatocytes from control and regenerating livers showed weak immunoreactivity for TGF-beta1. In contrast, the cytoplasm of hepatocytes of nodules in 7th, 8th and 9th month and HCCs were intensely stained for TGF-beta1. Some sinusoidal cells showed immunoreactivity for TGF-beta1 in all experimental groups. In early phase of carcinogenesis, the cytoplasm of hepatocytes in liver of 12h, 1d and 3d showed transiently increased immunoreactivity for TGF-beta1 and The immunoreactivity decreased thereafter. TGF-beta1 mRNA was also detected in the neoplastic hepatocytes by in-situ hybridization. Although TGF-betarII expression was correlated with TGF-beta1 immunoreactivity during early phase of carcinogenesis, hepatocytes in most nodules in 7th, 8th, 9th month and carcinomas showed decreased or little immunoreactivity for TGF-betarII. Based on the above results, it is concluded that TGF-beta1 expression increases not only in precancerous nodules but also in HCCs and its increase seems to be correlated with decrease or loss of TGF-betarII expression although its mechanism remains unclear. Hepatocytes may be a major cellular source of TGF-beta1 during hepatocarcinogenesis.

Expression Pattern of DNA Mismatch Repair Genes in Tumors of Microsatellite Mutator Phenotype.: Jung Jin Kim, Myung Jin Baek, Nam Gyun Kim, Yun Hee Kim, Ji Eun Kim, Hoguen Kim, Chanil Park; Korean J Pathol. 2000;34(9):609-614.

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Abstract PDF: Microsatellite mutator phenotype (MMP) tumors were reported in a subset of gastrointestinal carcinomas. The molecular pathogenesis of MMP tumors shows defects in the DNA mismatch repair genes, and also many germline and somatic mutations were reported in the MMP tumors. However, the detection of genetic defects in the MMP tumors is very difficult, mainly because many genes are included in the DNA mismatch repair genes. This study was undertaken to determine the best strategy for detecting defects in the DNA mismatch repair genes in gastrointestinal carcinomas. One of the effective ways for detecting defects in DNA mismatch repair genes is to screen the MMP tumors and evaluate the products of DNA mismatch repair genes by performing the multiplex RT-PCR method. We have screened the MMP tumors by using 5 microsatellite markers in the 12 cancer cell lines, 120 colon carcinomas and 99 gastric carcinomas and found 6 MMP cell lines, 10 MMP colon cancers, and 9 MMP gastric carcinomas. In addition, we evaluated 6 DNA mismatch repair gene products (hMSH2, hMSH3, hMSH6, hMLH1, hPMS1 and hPMS2) by multiplex RT-PCR analysis and found decreased expression of the DNA mismatch repair genes in 5 (hMSH6 in DLD-1 and HCT-15; hMSH2 in LoVo; hMLH1 and hMSH3 in HCT-116; hMLH1 in SNU-638) out of 6 MMP cell lines. We also found a decreased expression of hMLH1 in 3 out of 10 MMP colon carcinomas, and in 6 out of 9 MMP gastric carcinomas. Our results indicate that the expression analysis of the DNA mismatch repair genes by multiplex RT-PCR method can reduce the number of genes subjected to mutational analysis and is convenient for screening the responsible DNA mismatch repair genes.

Genomic Imbalances in Ependymoma by Degenerate Oligonucleotide Primed PCR-Comparative Genomic Hybridization.: Sung Hye Park, Gi Jin Kim, Min Kyung Kim, Hanseong Kim, Yoen Lim Suh, Sun Hwa Park; Korean J Pathol. 2004;38(3):133-137.

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Abstract PDF: BACKGROUND
The most consistent chromosomal abnormality in ependymomas, is loss of 22q (17-75%) and gain of 1q (0-50%). However, significance of this abnormality is uncertain.
METHODS
Genomic imbalances in 27 Korean ependymomas, including 21 low grade ependymomas, 4 anaplastic and 2 myxopapillary ependymomas, were analyzed by degenerate oligonucleotide primed-PCR-comparative genomic hybridization.
RESULTS
Common gains were found in 17 (63%), 20q (59%), 9q34 (41%), 15q24-qter (33%), 11q13 (30%), 12q23 (26%), 7q23-qter (26%), 16q23-qter (30%), 19 (26%), and 1q32-qter (22%). DNA amplification was identified in 12 tumors (44%). Chromosomal loss was a less common occurrence in our study, but was found in 13q (26%), 6q (19%), and 3 (11%).
CONCLUSION
The recurrent gains or losses of the chromosomal regions which were identified in this study provide candidate regions that may be involved in the development and progression of ependymomas.

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