Journal of Pathology and Translational Medicine

Volume 35(2); April 2001

Original Articles

mRNA is Synthesized Mainly at the Phase between the Euchromatin and Heterochromatin: Proposal of a Phase Theory.: Mi Sook Kim, Sang Woo Juhng; Korean J Pathol. 2001;35(2):93-97.

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Abstract PDF: BACKGROUND
Malignant cell nuclei, in general, have increased amounts of heterochromatin and decreased electron densities of euchromatin, making the chromatin pattern coarser than that of benign cell nuclei. The chromatin pattern in benign and malignant cells, however, is barely explained in terms of molecular structure. In this study, the chromatin pattern of metaplastic and carcinomatous squamous cells of the uterine cervix was correlated with transcriptional activity by ultrastructural autoradiography. METHODS: Punch-biopsied tissues were cultured with 3H-uridine for 5 minutes and processed for electron microscopy. Thin sections of the tissues on nickel grids were covered with photosensitive emulsion and kept cold in a dark room for 10 to 16 weeks. After development and staining, the tissues were observed by electron microscopy.
RESULTS
The nuclei of the metaplastic squamous cells consisted mostly of euchromatin. A few silver grains were observed, mainly at the periphery of the nuclei. The nuclei of the carcinomatous cells had increased amounts of heterochromatin along the nuclear membrane, and also in the euchromatin area. Silver grains were observed mainly at the boundary between the heterochromatin and euchromatin.
CONCLUSION
These findings suggest that an increased amount of heterochromatin in carcinomatous cells results in an increase of the boundary area between the heterochromatin and euchromatin, an area which may be a transcriptionally active site.

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.

Microsatellite Instability in Colorectal Carcinomas.: Hee Jeong Cha, Dong Kyun Woo, Sun Hee Kim, Yong ll Kim, Jae Gahb Park, Woo Ho Kim; Korean J Pathol. 2001;35(2):111-114.

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Abstract PDF: BACKGROUND
Microsatellite instability (MSI), which is caused by a deficient mismatch repair system, is seen in most of the hereditary non-polyposis colon cancers and a portion of sporadic colorectal cancers.
METHODS
Two hundreds forty-six consecutive sporadic colorectal cancer patients were analyzed for MSI using an ABI 377 automatic sequencer and fluorescent dye-labelled primers (BAT-25 and BAT-26).
RESULTS
The overall incidence of MSI in studied cases was 9.8% (24/246). This incidence is lower than most of the reported incidences in western countries. The incidence of MSI tumors in the proximal colon was 29.6%, while that of the distal colon was only 4.2% (p<0.001). MSI in sporadic colorectal cancers was more prevalent in poorly differentiated adenocarcinoma. In contrast to western countries, mucinous carcinoma did not show higher incidence of MSI.
CONCLUSION
The results suggest that MSI frequently occurs in cancers of the proximal colon and in tumors with poorly differentiated histology.

The Expression of Transforming Growth Factor-1 and Its Signaling Receptors in Human Colorectal Carcinoma.: Gyeong Seon Kim, Joo Heon Kim, Woo Sung Moon, Myoung Ja Chung, Dong Geun Lee, Myoung Jae Kang; Korean J Pathol. 2001;35(2):115-122.

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Abstract PDF: BACKGROUND
Resistance to the potent growth inhibitory effects of TGF- (transforming growth factor-) is a characteristic of many malignancies. TGF- insensitivity has been attributed to alterations in the number and function of the TGF- receptors as well as disturbances of downstream signal transduction. The aim of this study was to examine the expression of TGF-1 and its receptors in human colorectal cancer tissue and determine its relationship with cancer growth and with prognostic factors.
METHODS
Immunohistochemical staining of TGF-1, TGF-RI, and TGF-RII was performed on 20 human colorectal adenomas, 30 carcinomas and 10 normal mucosas as a control.
RESULTS
The staining indices of TGF-1, TGF-RI, and TGF-RII increased in adenomas and carcinomas compared with normal mucosas and adenomas, respectively. In adenomas the staining index of TGF-1 significantly increased with the severity of atypism. The staining index of TGF-RII increased in the carcinomas in the right colon and rectum, compared with those in the left colon.
CONCLUSION
The enhanced expression of TGF-1, TGF-RI and II in the colorectal carcinoma suggests an important role of colorectal carcinogenesis and tumor progression.

Pathological Analysis of 62 Liver Biopsy Cases with Hepatocellular Cholestasis: Drug and Toxin Induced Liver Injury.: Min Sun Cho, Young Nyun Park, Myeong Jin Kim, Kwang Jo Chae, Chanil Park; Korean J Pathol. 2001;35(2):123-128.

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Abstract PDF: BACKGROUND
Hepatocellular cholestasis denotes the alteration of bile secretion by hepatocytes. The causes, degree of hepatocyte injury and concomitant bile duct loss are considered to influence the clinical course.
METHODS
The causes and pathological features of hepatocellular cholestasis were analyzed in 62 cases of liver biopsies; and the causes of primary biliary cirrhosis, primary sclerosing cholangitis, and biliary obstruction were not included.
RESULTS
The mean age of the patients was 42.2 years, and the ratio of male to female was 1.8:1. Fifty-eight cases (94%) showed cholestatic hepatitis, and 4 cases (6%) showed pure cholestasis without hepatitis activity. The majority of the cases (52 cases, 84%), including 19 cases of herbal medicine, was related to drugs. Loss of bile duct was found in 12 cases (19%), which were all cases of chronic cholestasis. All of them had drug histories, including 9 cases of herbal medicine. Clinical follow-up was performed in 9 out of the 12 cases with bile duct loss, and all of them showed elevated total bilirubin and/or alkaline phosphatase levels for more than 6 months.
CONCLUSION
Drugs are the major cause of hepatocellular cholestatic hepatitis/cholestasis; and information about drugs, including herbal medicines, should be considered for proper evaluation of liver biopsy with hepatocellular cholestasis. Bile duct loss should be evaluated in the cases of chronic hepatocellular cholestasis, especially in drug induced cases.

A Study on the Expression of Proliferating Cell Nuclear Antigen and Apoptosis of the Hepatocellular Carcinoma in Human and Hepatitis B Virus X Transgenic Mice.: Hyung Bae Moon, Dae Yeul Yu, Hyung Ryun Yoo, Byung Joon So, Kwon Mook Chae, Haak Cheol Kim, Ki Jung Yun, Won Cheol Han, Hyang Jeong Jo, Bo Yong Kim; Korean J Pathol. 2001;35(2):129-136.

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Abstract PDF: BACKGROUND
This experiment was designed to study the cell kinetics of hepatocellular carcinoma (HCC) in both hepatitis B virus X (HBx) transgenic mice and humans.
METHODS
The immunohistochemical stain of proliferating cell nuclear antigen (PCNA) and TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay of apoptosis were used on formalin fixed-paraffin embedded tissues.
RESULTS
PCNA labeling indices (PCNA-LI) in the liver of HBx transgenic mice were markedly increased in HCC (11.3%) compare to the dysplastic areas (1.3%) and in the liver of non-transgenic littermates (0.1%). There was no significant difference of PCNA-LI in the dysplastic areas between HCC developed mice and non-HCC developed mice. Apoptosis labeling indices (Apoptosis-LI) in both the dysplastic areas and HCC of HBx transgenic mice were similar to those of non-transgenic littermates. PCNA-LI was markedly increased in human HCC (28.9%) compare to the background of HCC (2.9%) and the control liver (2.9%). Apoptosis-LI was decreased in human HCC (0.3%) compare to the background of HCC (0.4%) and the control liver (1.0%). Conclusion : There is a marked increase of cell proliferating activity in human HCC and in HCC of HBx transgenic mice, and there is a decrease of apoptosis in human HCC, but not in HCC of HBx transgenic mice.

Study of Microsatellite Alterations of 3p and 11q Chromosomes in Uterine Cervical Adenocarcinoma.: Eung Seok Lee, Hye Jin Jeong, Hee Jeoung Kim, Insun Kim; Korean J Pathol. 2001;35(2):137-143.

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Abstract PDF: BACKGROUND
Uterine cervical cancer is the most prevalent cancer in Korean women, and the incidence of adenocarcinoma has been increasing. Loss of heterozygosity (LOH) analysis is used to identify regions which harbor a putative tumor suppressor gene.
METHODS
DNA was extracted from the microdissected normal and malignant lesions of 34 uterine cervical adenocarcinomas, 2 adenosquamous cell carcinomas, 13 squamous cell carcinomas, and 10 endometrial adenocarcinomas. LOH and microsatellite instability (MSI) analysis were performed using microsatellite markers, D3S4103 (3p14.2), D3S1284 (3p12), D3S1289 (3p21.2-21.1), D3S1307 (3p25-ter), THRB (3p22-24.1), and D11S35 (11q22). The expression of Fhit protein was compared with the genetic abnormalities.
RESULTS
Microsatellite alterations at 3p were detected in 37% of cervical adenocarcinomas, 16% of squamous cell carcinomas, and 43% of endometrial adenocarcinomas. The alterations of 11q were found in 17% of cervical adenocarcinomas. Microsatellite alterations of D3S1307 and D11S35 were detected in uterine cervical adenocarcinomas with high frequency. The frequency of FHIT protein loss is higher in the cervical squamous cell carcinoma than in cervical and endometrial adenocarcinomas.
CONCLUSION
Tumor suppressor gene of uterine cervical adenocarcinoma may be located in 3p25-ter and 11q22.

Expression of p16 and Rb in 9,10-Dimethyl-1,2-Benzanthracene Induced Rat Ovarian Carcinogenesis.: Ki Kwon Kim, Dong Hun Kim; Korean J Pathol. 2001;35(2):144-150.

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Abstract PDF: BACKGROUND
In order to investigate the roles of p16 and Rb, their expression was evaluated in 9,10-dimethyl-1,2-benzanthracene (DMBA)-induced ovarian cancers of rats.
METHODS
DMBA-coated silk was inserted into both ovaries of 20 9-week-old Sprague-Dawley rats. The experimental period lasted 20 weeks. The tumor histology was classified and the expression of p16 and Rb in the ovarian tumors was analyzed by immunohistochemistry and Western blot.
RESULTS
The p16 and Rb labeling index was significantly lower in the ovarian cancers than the normal ovarian surface epithelium of a rat. There were no differences among the cancer types. In Western blot analysis, the expressions of p16 and Rb in ovarian cancers were lower than those in normal ovarian tissue. No correlation was present between p16 and Rb.
CONCLUSION
The abnormal expression of p16 and Rb occurs in DMBA-induced rat ovarian cancer and might be involved in carcinogenesis.

Expression of Cyclin D1, Cyclin E, p21Cip1 and p27Kip1 in Chemically Induced Rat Mammary Tumor Treated with Tamoxifen and Transforming Growth Factor-1.: Tae Jung Jang, Jae Hum Park, Mee Yon Cho, Ki Kwon Kim, Jung Ran Kim; Korean J Pathol. 2001;35(2):151-157.

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Abstract PDF: BACKGROUND
Tamoxifen (TAM) inhibits the action of estrogen by binding to estrogen receptors, and also has non-estrogen receptor mediated cytostatic activities. Transforming growth factor-1 (TGF-1) inhibits the proliferation of many other cell types, such as epithelial, hematopoietic and endothelial cells.
METHODS
We investigated the effects of tamoxifen on the growth of 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumors and the expression of cyclin D1, cyclin E, p21Cip1, and p27Kip1 by performing immunohistochemistry and Western blot analysis, and studied whether TGF-1 injection amplified the effects of TAM. When tumor size reached between 10-15 mm in the largest dimension, the rats were divided into 3 groups: DMBA-control group (n=12), DMBA-TAM group (n=14) and DMBA-TAM plus TGF-1 group (n=5).
RESULTS
The consecutive administration of TAM markedly decreased the tumor development compared with the DMBA-control group. The DMBA-TAM and DMBA-TAM plus TGF-1 groups showed decreased expression of bromodexoyuridine, cyclin D1, cyclin E, and p21Cip1 when compared with those of the DMBA-control group. On the other hand, the labeling index of p27Kip1 was higher in the DMBA-TAM plus TGF-1 group than in the DMBA-control group.
CONCLUSION
TAM suppresses tumor development, which may be associated with down-expression of cyclin D1 and cyclin E, and overexpression of p27Kip1, and addition of TGF-1 does not influence tumor development treated by TAM.

Ethnic Differences of the p53 Genetic Alteration in Cutaneous Malignant Melanoma.: Won Sang Park, Eun Young Na, Sang Kyu Lee, Sug Hyung Lee, Su Young Kim, Seok Jin Kang, Kye Yong Song, Suk Woo Nam, Nam Jin Yoo, Jung Young Lee; Korean J Pathol. 2001;35(2):158-164.

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Abstract PDF: BACKGROUND
There are significant differences in the clincopathologic pattern including the incidence, favor site, and histopathologic type between cutaneous malignant melanomas arising from whites, asians and blacks. These differences might suggest that there is a racial difference in the molecular tumorigenesis mechanism of malignant melanoma.
METHODS
To determine the ethnic differences in tumorigenesis of malignant melanoma, we performed loss of heterozygosity (LOH) and sequencing analyses of the p53 gene in cutaneous malignant melanomas arising from 22 white American, 30 Korean and 15 black African patients.
RESULTS
The frequency of LOH of the p53 gene is only 12.5% in white American patients, but the frequency is significantly higher in Korean (42.1%) and black African (61.5%) patients. We also detected 17 mutations (nonsense: 1, missense: 16) of the p53 gene in the cutaneous malignant melanomas of Koreans and black Africans, but none in those of white Americans: among the 16 missense mutations, 10 mutations were C:G to T:A transitional mutations. Of these, we also detected one GG (CC) to AA (TT) tandem mutation at the pyrimidine sequence.
CONCLUSION
These results strongly suggest that there might be a racial difference in molecular carcinogenesis mechanisms among the cutaneous malignant melanomas occurring in white American, Korean and black African patients. But the role of the p53 genetic alteration in the genesis of melanomas in Korean and black African patients is subject to further evaluation.

Case Reports

Pulmonary Aspergillus niger Fungus Ball Associated with Calcium Oxalate Crystal: A Case Report.: Eunhee Lee, Heejung Kim, Eundeok Chang, Kyo Young Lee, Byung Kee Kim; Korean J Pathol. 2001;35(2):165-167.

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Abstract PDF: Pulmonary aspergillomas are considered to represent saprophytic growth of hyphae within the preexisting lung cavity. Causative Aspergillus species are mostly composed of A. fumigatus and A. flavus and, occasionally, A. niger. It is difficult to identify these species using histopathologic evaluation without a culture of sputum and bronchoalveolar lavage. We recently experienced a case of pulmonary Aspergillus niger infection associated with characteristic calcium oxalate crystal formation. A large cavity contained hyphae with a typical conidial head, conidiophore, and conidia. Characteristic birefrigent calcium oxalate crystals were demonstrated in the cavity wall.

Polyomavirus Renal Infection Confirmed by Electron Microscopy in a Patient with Acquired Immunodeficiency Syndrome: An Autopsy Case Report.: Na Rae Kim, Byoung Kwon Kim, Je G Chi; Korean J Pathol. 2001;35(2):168-171.

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Abstract PDF: Polyomavirus infection commonly occurs in childhood and adolescence, remaining in a latent status and reactivated in an immunocompromised status. We report herein an autopsy case of HIV-positive 41-year-old male, who succumbed to disseminated Kaposi sarcoma and cytomegalovirus infection involving the gastrointestinal tract, lung and brain. The involved kidney showed minimal inflammatory infiltrates and tubular injury: the nuclei of tubular epithelial cells were markedly enlarged with central clearing and peripheral chromatin margination or bore basophilic nuclear inclusions. Inclusion-bearing tubular epithelial cells were negative for the viral immunostains including herpes simplex virus, Epstein-Barr virus and adenovirus. Electron microscopy disclosed 42 nm intranuclear viral particles compatible with the BK polyomavirus. The viral particles were icosahedral in paracrystalline array and nonenveloped.

Atypical (Bizarre) Leiomyoma of the Prostate: A Case Report.: Sung Rim Kim, Sang Yong Song, Geunghwan Ahn, Han Yong Choi; Korean J Pathol. 2001;35(2):172-175.

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Abstract PDF: Atypical (bizarre) leiomyoma of the prostate is a very rare neoplasm. Five cases have been reported in English medical literature. A 60-year-old Korean man with a history of prostatism and slightly elevated serum prostate specific antigen was presented. Microscopically, the transurethral resection specimen consisted of a proliferation of hypercellular spindle cells with intersecting bundles. The nuclei of the tumor cells showed marked pleomorphism and hyperchromasia with occasional multinucleated giant cells. Mitoses were seen in areas of up to 2 per 10 high power fields, but there was no evidence of atypical ones. The tumor cells were immunoreactive against anti-smooth muscle actin and desmin antibodies. The proliferative index (10.0%) of the atypical leiomyoma lay between that of a benign smooth muscle and that of a leiomyosarcoma of the prostate. Flow cytometry showed a diploid pattern with an elevated S phase fraction. To the best of our knowledge, this is the first demonstration of atypical leiomyoma of the prostate in a Korean man.

Choroid Plexus Carcinoma: A Report of Two Cases.: Young Ran Shim, Mi Jin Gu, Dong Sug Kim, Oh Lyong Kim, Woo Mok Byun, Yong Jin Kim; Korean J Pathol. 2001;35(2):176-179.

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Abstract PDF: Choroid plexus carcinoma (CPC), a frankly malignant epithelial neoplasm derived from choroid plexus epithelium, is a rare tumor with a predilection for infants and children. It may be difficult to histologically differentiate it from choroid plexus papilloma, anaplastic ependymoma, medulloblastoma, germ cell tumors, and metastatic carcinoma. We examined two cases of CPC. One is a 12-month-old boy, and the other is a 13-month-old boy. Both patients present lateral ventricular masses with extensive hydrocephalus. Histologically, both tumors show papillary growth in most area, and focal solid growth. The tumor cells show marked nuclear pleomorphism and frequent mitoses on squash and hematoxylin-eosin slides. Immunohistochemically, both tumors are positive for cytokeratin, vimentin, and S-100 protein; but they are negative for glial fibrillary acidic protein, -fetoprotein, and placental alkaline phosphatase. Both tumors show diffuse and strong positivity for p53. The MIB-1 labelling index is 23.6% and 15.82%, respectively. We report two cases of typical CPC, and we briefly discuss differential diagnosis with review of literatures.

Metastatic Alveolar Soft Part Sarcoma Presented with a Dura Mass: A Case Report.: Sun Young Kwon, Hyun Chang Joo, Kun Young Kwon, Sang Sook Lee, Chul Ho Sohn, Man Bin Yim, Sang Pyo Kim; Korean J Pathol. 2001;35(2):180-183.

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Abstract PDF: Alveolar soft part sarcoma, accounting for less than 1% of all soft tissue sarcomas, is known for late metastases to the lung, bone and brain. We have examined a case of metastatic alveolar soft part sarcoma to the brain in a 34-year-old woman. Computed tomography of the brain revealed a well enhancing dura based mass of the left temporal area. The patient had a previous history of surgery for alveolar soft part sarcoma from the right thigh at the age of 24 years. She underwent total removal of the dural mass under the clinical impression of the meningioma. Grossly, a well-circumscribed, pale tan and solid mass was 3.4 x 3.2 x 2.4 cm and easily detached from the attached dura. Histologically, the tumor showed typical features of alveolar soft part sarcoma. Immunohistochemically, tumor cells were positive for muscle related antibodies. Ultrastructural examination showed rod-shaped membrane bound crystals and electron-dense granules in the cytoplasm of tumor cells.

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