Journal of Pathology and Translational Medicine

Volume 31(8); August 1997

Original Articles

Establishment and Characterization of a Small Cell Lung Cancer Cell Line(JePa-1).: Mi Ja Lee, Ho Jong Jeon, Jong Hoon Chung; Korean J Pathol. 1997;31(8):695-710.

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Abstract PDF: Lung cancer is the most common malignant tumor worldwide and its incidence continues to rise each year. Recent development of molecular biologic method has led to advances in determining the etiologic factors of lung cancer and the establishment of cell lines has provided a lot of information on the through chemosensitivity, radiation biology studies, cytogenetics, and molecular biologic studies, which permits improved treatment for lung cancer. We established a small cell lung cancer cell line, designated JePa-1, obtained from malignant pericardial effusion of small cell lung cancer patient and characterized its morphologic and molecular biologic features. the JePa-1 cell line grew relatively slowly (doubling time 45hrs) as very loosely adherent floating aggregates growing in small clumps with distinct cell outlines and intertwined cords. Also JePa-1 cell line secreted antidiuretic hormones. Electronmicroscopic examination revealed that JePa-1 cell line and xenografts contained electron dense core granules, characteristic of being of neuroendocrine origin. To investigate the tumorigenic capacity, the JePa-1 cell line was injected into SCID and nude mice. Tumors taken from xenografts were observed in 3 out of 4 of the SCID mice and 2 out of 4 of the nude mice. The histologic characteristics of the xenografts were similar to those of the cell line and the original cytologic finding of the pericardial fluid, suggesting small cell carcinoma. The results of immunohistochemical markers showed reactivity for Rb protein, c-myc, TGF-alpha, TGF-beta , EGFR, keratin, NSE, chromogranin, and EMA. The DNA ploidy and the index of the JePa-1 cells was tetraploid and 2.13, respectively. The positive rate for the Rb, c-myc and K-ras proteins of the JePa-1 cell line were 98.9%, 99.3%, and 99.7% respectively as determined by flow cytometry. Cytogenetic analysis using the G-banding technique showed 65 chromosomes with various numerical and structural abnormalities. On examination of the expression of TGF-alpha, TGF-beta , and EGFR by PCR, only the EGFR was positive Through the establishment of JePa-1 cell line, we report in this paper the characterization of a small cell lung cancer such as morphologic and immunocytochemical features, growth characteristics in culture, hormone production, expression of oncoprotein and several growth factors, tumorigenicity, chromosomal abnormalities, and DNA ploidy and index. The JePa-1 cell line will be valuable in vitro studies for the etiology, treatment and the prognostic factors in small cell lung cancer.

Experimental Study on Shark Liver Oil-Induced Lipoid Pneumonia in Rats.: Mee Soo Chang, Eui Keun Ham; Korean J Pathol. 1997;31(8):711-722.

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Abstract PDF: The purpose of this experiment is to evaluate the histopathologic findings of shark liver oil-induced lipoid pneumonia, and to determine whether shark liver oil is absorbed through lymphatics and the venous system or not. A single intratracheal administration of shark liver oil (0.6 ml/kg of B.W.) was given to Sprague-Dawley rats. They were then sacrificed sequentially from 1 hour to 12 weeks after injection. We investigated the chest radiographic findings, the serum total lipid concentration of blood obtained by cardiac puncture, lipid-laden alveolar macrophage index of the bronchoalveolar lavage fluid, and the histopathology of tracheobronchial lymph nodes and the lung (Oil red O stain & H&E stain). Chest radiographs showed no specific findings; ill-defined hazy, linear, small patch radioopacity, air space consolidation or collapse. Thirty-six percent of the experimental rats revealed normal findings. Within the lung, the shark liver oil appeared either as highly emulsified fine granules in the cytoplasm of the alveolar macrophage or as free, round oil masses. The area of the lung accumulated with lipid material was maximized 1 week after injection, and then decreased thereafter. The tissue reactions were cuboidal metaplasia of the alveolar lining, widening and lymphocytic infiltration of the alveolar septa and granuloma formation (3% of experimental rats) as a reaction to a foreign body. There were also lung abscesses due to superimposed bacterial infection (5% of experimental rats). With time after the injection of the oil, the serum total lipid tended to increase and the intracellular lipid of the alveolar macrophages in the bronchoalveolar lavage fluid tended to decrease. In summary, the histopathologic findings of the lung in the experimental lipoid pneumonia were interstitial chronic inflammation and granulomas with the presence of lipoid material in the lung parenchyma, and shark liver oil appeared to be absorbed in the blood and the lymph, then metabolized.

Expression of Epstein-Barr Virus Gene Products, Bcl-2 and p53 Proteins in Nasopharyngeal Carcinomas.: Sun Hee Yoon, Kang Suek Suh, Chang Hun Lee; Korean J Pathol. 1997;31(8):723-734.

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Abstract PDF: The authors studied EBV genome expression in 40 conventionally processed samples of nasopharyngeal carcinomas (NPC), using in situ hybridization for EBERs and immunohistochemistry for LMP, Bcl-2 and p53 proteins. The NPCs consisted of 6 keratinizing squamous cell carcinomas (KSCs), 13 nonkeratinizing carcinomas (NKCs) and 21 undifferentiated carcinomas (UCs). The results were summarized as follows: 1) EBERs were expressed in 80.0% of all the NPCs (32/40). As for the subtypes, they were detected in 92.3% of NKCs (12/13), in 90.5% of the UCs (19/21), and in 16.7% of the KSCs (1/6). In positive cases, the nuclei of tumor cells displayed uniformly strong staining. 2) LMP was expressed in 10.0% of all the NPCs (4/40), all of which were UC. The LMP expression in the UCs was not correlated to the expression of EBERs, Bcl-2 and p53 proteins. 3) Bcl-2 protein was detected in 85.0% of all the NPCs (34/40). As for the subtypes, they were detected in 92.3% of the NKCs (12/13), in 90.5% of the UCs (19/21), and in 50.0% of the KSCs (3/6). 4) p53 protein was detected in 75.0% of all the NPCs (30/40). As for the subtypes, they were detected in 81.0% of the UCs (17/21), in 69.2% of the NKCs (9/13), and in 66.7% of the KSCs (4/6). 5) In the NPCs the expression of EBER showed a significantly positive correlation with that of p53 or Bcl-2 protein. The above results indicate that the association of EBV with NPC is chiefly with poorly differentiated and undifferentiated carcinomas. Additionally, carcinomas commonly display widespread, strong immunoreactivity of Bcl-2 and p53 proteins over tumor cells. In conclusion, these observations indicate that the EBV-association in NPC appears to contribute to the overexpression of tumor-related genes during carcinogenesis.

Tumor Angiogenesis and Cathepsin-D Expression in Invasive Ductal Carcinoma of the Breast.: Young Gyung Bae, Dae Hong Suh, Dong Sug Kim, Soo Jung Lee; Korean J Pathol. 1997;31(8):735-744.

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Abstract PDF: This study was conducted to assess the prognostic value of tumor angiogenesis and Cathepsin-D in breast carcinoma, by correlating them with other clinicopathologic prognostic factors. In order to estimate microvessels within the tumor, an immunohistochemical method using monoclonal antibodies for factor VIII-related antigens (DAKO-vWf, F8/86) was used, and they were counted (perx200 field) in the most active areas of neovascularization. For the expression of Cathepsin-D, an immunohistochemical method using monoclonal antibodies (Novocastra, NCL-CDm) was performed. The microvessel count ranged from 8 to 346 per x200 field and the mean (+/-SD) was 72.46+/-54.96. The microvessel count was correlated with the estrogen receptor status, and it was also correlated with the tumor size when it was graded into four groups (1-33, 34-67, 68-100, >100), but was not correlated with other clinicopathologic parameters. Cathepsin-D was expressed in 40% (46/115) of the cases, but it was statistically correlated with the tumor size only. In conclusion, the expression of Cathepsin D and the degree of angiogenesis in breast cancer showed a correlation with the tumor size only. Therefore, they do not appear to be good prognostic parameters, according to the present study.

Immunohistochemical Study of E-cadherin Expression in Gastric Adenocarcinomas.: Jee Yeon Kim, Mee Young Sol, Sun Kyung Lee; Korean J Pathol. 1997;31(8):745-753.

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Abstract PDF: E-cadherin (ECD) is a Ca++ -dependent adhesion molecule which plays a major role in the maintenance of intercellular adhesion in epithelial tissues. The expression pattern of ECD in 77 surgically resected gastric adenocarcinomas was examined by immunohistochemistry, using a rat monoclonal antibody raised against murine E-cadherin (DECAM-1). ECD was strongly expressed uniformly at cell to cell borders in normal gastric epithelium without exception. But, various staining patterns were observed in the cancer tissues. The frequency of tumors with preserved ECD expression (Pre-type) and reduced ECD expression (Rd-type) was 44% and 56%, respectively. Using Lauren's classification, the high frequency of the Pre-type expression in adenocarcinoma of the intestinal type was significantly higher than that in adenocarcinoma of the diffuse type (p<0.05). But, no significant correlation between the ECD expression and the gross type, invasion depth, growth pattern or metastasis was observed. These results suggest that ECD might play a key role in the morphogenesis of gastric adenocarcinoma.

Pathologic Diagnosis of Intestinal Tuberculosis in Endoscopic Biopsied Material.: Kyoung Mee Kim, An Hi Lee, Kyu Yong Choi, Se Jeong Oh, Sang In Shim; Korean J Pathol. 1997;31(8):754-764.

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Abstract PDF: The clinicopathologic features and the comparative analysis of diagnostic methods in 42 patients having intestinal tuberculosis were studied. In all the cases, clinical and colonoscopic diagnosis was confirmed by histological examination. Abdominal pain was the most common symptom (54%). Twenty nine patients had active pulmonary tuberculosis which was confirmed by a chest X-ray, or an AFB smear and a culture of sputum. A transverse ulcer with surrounding hypertrophic mucosa and multiple erosion was the usual colonoscopic findings. The granulomas were usually located in the just upper and lower portion of muscularis mucosa. The direct smear and culture of the fresh biopsy material showed AFB in 11 (32.4%) and 12 cases (36.4%) respectively. Ziehl-Neelsen staining in serially sectioned slides from formalin-fixed, paraffin- embedded tissue revealed AFB in 15 cases (35.7%). An immunohistochemical stain for Mycobacterium bovis was done in all cases and 13 cases were positive (31%). A polymerase chain reaction (PCR) was done and showed positivity in 4 out of 20 cases of fresh biospy material and 12 out of 40 cases in paraffin embedded tissue. For the conclusive diagnosis of intestinal tuberculosis, a Ziehl-Neelsen stain is the most sensitive, fast, and cost-effective method. The diagnostic accuracy will be increased when other diagnostic methods such as tissue culture and PCR are coupled with this simple staining method.

An Anion Site Change of the Glomerular Basement Membrane on Various Glomerular Diseases.: Yu Na Kang, Kwan Kyu Park, Seung Pil Kim, Sung Bae Park, Hyun Chul Kim, Eun Sook Chang, In Soo Suh; Korean J Pathol. 1997;31(8):765-772.

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Abstract PDF: We studied the ultrastructural alteration of glomerular anionic sites in 6 patients with minimal change nephrotic syndrome, 5 patients with membranous glomerulonephritis, 4 patients with focal segmental glomerulosclerosis, and 4 patients with IgA nephropathy by staining with polyethyleneimine (PEI) as a cationic probe. The control study was examined by using a nephrectomy specimen of non-glomerular disease which had no proteinuria. This method seems to selectively stain heparan sulphate in the basement membranes and has been widely used to evaluate changes in basement membrane charge in various human diseases as well as in experimental studies. The anionic sites in the lamina rara interna and lamina densa of normal glomerular basement membrane were always less numerous and less regularly distributed than those in the lamina rara externa. Characteristic common findings in these glomeruli showed a marked decrease of glomerular anionic sites in the regions with immune-complex deposits and normal distribution in the regions with focally those being absorbed and newly forming glomerular basement membrane. They were not detected in the gap of the basement membrane and on the area of the detached overlying epithelium using the PEI method. But the foot process fusion of epithelial cells seems not to influence the loss of anionic sites on the glomerular basement membrane.

Relationship between Proliferative Activity and Expression of HBcAg and p53 Protein in Hepatocellular Carcinoma and Surrounding Nontumorous Liver.: So Ya Paik, Ho Guen Kim, Chan Il Park; Korean J Pathol. 1997;31(8):773-781.

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Abstract PDF: In an attempt to discover the factors contributing to the increased proliferative activity in hepatocytes and subsequent development of HCC, the proliferative activity of hepatocytes was compared with the size of regenerative nodules and HBcAg expression status in the surrounding nontumorous liver of 45 surgically resected hepatocellular carcinomas, including 34 HBV related ones. In the tumor, the difference in proliferative activity and the histological grade was analyzed in terms of p53 gene alteration. The proliferative activity was assessed by immunohistochemical methods using Ki-67 monoclonal antibody. HBcAg expression in the surrounding nontumorous liver correlated with both the inflammatory and proliferative activity of hepatocytes (p<0.05). p53 overexpression was associated with high proliferative activity and aggressive phenotype of tumor. No correlation was observed between the proliferative activity of hepatocytes and the size of regenerative nodules in cirrhosis (p>0.05). p53 overexpression was not evident in surrounding nontumorous liver including cirrhosis. In conclusion, the above results are in line with the view that hepatic carcinogenesis is a mutistep, progressive process. In the initial stage, chronic cellular injury incurred by immumologic reaction against HBcAg seems to play a pivotal role in increased cellular regeneration. However, once transformation of hepatocytes occur the major contributor to tumor growth seems to be alteration in p53 tumor suppresor gene.

Case Report

Clear Cell Meningioma.: Hee Jeung Cha, Soo Kee Min, Joon Mee Kim, Young Chae Chu; Korean J Pathol. 1997;31(8):782-787.

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Abstract PDF: Clear cell meningioma is a recently recognized morphologically unique entity. It shows no sex predilection, affects primarily the lumbar region, and the cerebellopontine angle. Despite its benign appearance, it may be aggressive, particularly in intracranial cases. All lesions are moderately cellular, with the exception of stromal hyalinization. The tumor consists largely of a sheet- like or somewhat lobular pattern of polygonal cells, the cytoplasm of which is clear. No close association is noted between the recurrence or the clinical outcome and factors such as mitotic activity, the PCNA index, and the DNA ploidy status. But the MIB-1 proliferation index is appreciably higher in recurrent tumors. We experienced a case of clear-cell meningioma showing a characteristic histologic finding. A 39-year-old man was admitted due to the recent onset of right-sided, facial-nerve palsy, left hemiparesis and general weakness. A CT scan of the head showed a well defined mass in the petroclival area. After surgical resection, the patient was in good condition, but 1 year later symptoms recurred. A CT scan of the head showed a huge, recurrent petroclival tumor with adhesion to the surrounding brain parenchyme.

Original Article

Composite Adenocarcinoma and Choriocarcinoma of the Sigmoid Colon with Hepatic Metastasis of the Choriocarcinomatous Component.: Young Ha Oh, Won Mee Lee, Kyung Sook Kim, Moon Hyang Park, Jung Dal Lee; Korean J Pathol. 1997;31(8):788-793.

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Abstract PDF: A rare case of hepatic metastasis with a choriocarcinomatous component from a composite adenocarcinoma and choriocarcinoma of the sigmoid colon in a 60-year-old man is reported. The hepatic metastasis displayed choriocarcinoma with extensive hemorrhagic necrosis. The tumor cells were poorly differentiated with scattered foci of bizzare syncytiotrophoblastic cells. Retrospective examination of the previous colonic carcinoma proved that the tumor was composed of two distinctive elements. One was a moderately well differentiated adenocarcinoma located in mucosa and submucosa. The other was a deep seated and undifferentiated carcinoma which was made up of hyperchromatic bizzare cells with syncytiotrophoblastic cells. There were transitional foci from adenocarcinoma to undifferentiated carcinoma with trophoblastic cells. Immunohistochemical staining showed beta-hCG expression in the undifferentiated cells of both the primary and the metastatic tumors. Implications for the possible origin and cause of tumor cell heterogeneity are briefly discussed.

Case Reports

Choriocarcinoma of the Colon.: Youn Mee Kim, Mee Youn Cho, Soon Won Hong, Soon Hee Jung; Korean J Pathol. 1997;31(8):794-797.

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Abstract PDF: Choriocarcinoma of the gastrointestinal tract is rare. Among them, that of the stomach is the most common. Six cases of choriocarcinoma of the colon were found in the review of the literature. All of these previously reported cases had multiple metastatic foci in the liver, lung, lymph nodes and the prognosis seemed to be very poor. Therefore we think that choriocarcinoma of the colon should be distinguished from conventional adenocarcinoma. A 66-year old female patient, described in this case, was operated on under the impression she was suffering from acute appendicitis. The resected ascending colon revealed extensive hemorrhagic necrosis and perforation with fibrous adhesion in the cecum. On the cut section, the mural tumorous thickening was not definite. Histologically, the tumor showed a focus of typical adenocarcinoma arising from glandular epithelial cells, which were transformed into highly anaplastic tumor cells. There were frequent vascular invasions of tumor cells, similar to syncytiotrophoblasts. In the immunohistochemical stains, both glandular and highly anaplastic tumor cells reacted with cytokeratin. The glandular cells were also reactive for carcinoembryonic antigen (CEA) and anaplastic tumor cells for human chorionic gonadotrophin (hCG). This is the first report of choriocarcinoma of the colon in Korea. We describe this case with a review of the literature.

Intracranial Fibro-Osseous Lesion: A case report and literature review.: Jae Weon Lim, Seung Cheol Lee, Byoung Yuk Yi, Yoon Kyung Sohn; Korean J Pathol. 1997;31(8):798-801.

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Abstract PDF: Intracranial fibro-osseous lesion, also reported as calcifying pseudoneoplasm of the neural axis, is an uncommon lesion of the central nervous system. Since the discovery of this entity by Rhodes and Davis in 1978, there have been a total of 21 cases reported in the literature. We encountered one such case in a 28 year old male, who presented with left hemiparesis for 1 year. By the MR images, a 1.5 cm sized round mass was found at right parietal lobe near motor cortex. The mass lesion enhanced well, homogenously and revealed clear, slightly irregular margin. Excisional biopsy of the mass was performed. Microscopically the lesion was composed of calcified fibrous tissue with an amorphous gray-blue, coarsely fibrillar to chondromyxoid nodular areas. Sparse spindle cells, immunohistochemically negative for GFAP, vimentin and S-100, were scattered within the amorphous material. Palisading spindle or polygonal cells were present at the more cellular periphery of the lesion, which were vimentin positive but S-100 negative. There was no evidence of the pilocytic astrocytes, Rosenthal fibers, or GFAP positive hypertrophic astrocytes. Intracranial fibro-osseous lesions are apparently slow-growing with generally excellent prognosis after wide excision. The etiology remains unclear, but most investigators favor a reactive rather than neoplastic process.

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