Journal of Pathology and Translational Medicine

Volume 32(5); May 1998

Original Articles

Identification of Differentially Expressed Genes Using RNA Fingerprinting in Cell after DNA Damage.: Jung Young Lee, Min Sun Shin, Seung Myung Dong, Eun Young Na, Su Young Kim, Sug Hyung Lee, Won Sang Park, Nam Jin Yoo; Korean J Pathol. 1998;32(5):321-327.

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Abstract: RNA fingerprinting using on arbitrary primed polymerase chain reaction (RAP-PCR) was carried out to identify differentially expressed genes in HL-60 cell after treatment of methylmethane sulfonate (MMS). Twenty differentially expressed PCR products were cloned and analyzed. We have successfully obtained eight partial cDNA sequences by TA cloning method. Among these, six cDNAs were up-regulated and two cDNAs were down-regulated after the MMS treatment. Of these six up-regulated cDNAs, 3 cDNAs were equivalent to known genes in the GenBank/EMBL databases with 98~100% homology searched by BLAST program: genomic DNA fragment containing CpGg island (clone 26h8), Human Rev interacting protein-1 (RIP-1), and human zinc finger protein-4 (HZF4). The sequences of the three remaining cDNA were entirely new genes, but we didn't try to identify a full cDNA sequence. Two clones called KIAA0060 and KIAA0065, were down-regulated in HL-60 cells after the MMS treatment. These findings suggest that the RNA fingerprinting method using RAP-PCR is an effective method which can identify and separate the differentially expressed cDNAs and that the isolated cDNAs might involve in regulation mechanism of apoptosis and/or cell cycle delay, especially a p53-independent pathway, in the cells after DNA damage. But the nature of cDNAs that we have isolated remains to be elucidated.

Synaptic Reorganization of Dentate Mossy Fibers and Expression of Calcium Binding Proteins in Hippocampal Sclerosis of Temporal Lobe Epilepsy.: Sang Pyo Kim, Seung Pil Kim, Seung Che Cho, Young Rok Cho, Ji Min Jeon, Yu Na Kang, Kun Young Kwon, Eun Sook Chang; Korean J Pathol. 1998;32(5):328-336.

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Abstract: This study was designed to identify expression of calcium-binding proteins and synaptic reorganizations of dentate mossy fibers in hippocampal sclerosis of human temporal lobe epilepsy. Hippocampal neuronal density was quantitively analyzed in temporal lobe epilepsy group (n=50) to investigate the degree of hippocampal sclerosis and it was compared with that of autopsy control (n=3). To verify the distribution of calcium-binding proteins in neurons of epileptic hippocampi, the parvalbumin (PV)-immunoreactive and calbindin-D28K (CB)-immunoreactive neurons were quantitively analyzed in each area of Ammon's horn by immunohistochemical stain. Also, to clarify synaptic reorganizations of the dentate mossy fibers, a part of each hippocampus was examined under light microscopy and transmission electron microscopy using Timm sulphide silver method. In epileptic hippocampi, severity of hippocampal sclerosis (HS) was graded four, which consisted of 3 cases with no HS, 6 mild HS, 12 moderate HS, and 29 severe HS. The hippocampal neuronal loss was most prominent in CA1, followed by CA4 and CA2. Expression of calcium-binding proteins was more prevalent in CA2 of all groups. The proportion of PV-immunoreactive neurons in CA1 and CA4 significantly increased in the moderate and severe HS group, whereas the proportion of CB-immunoreactive neurons did not correlated with the severity of HS. Timm granules were noted in inner molecular supragranular layer of dentate gyrus of epileptic hippocampi and they tended to increase in proportion along with the severity of hippocampal sclerosis. Transmission electron microscopy showed that supragranular Timm granules corresponded to synaptic terminals of mossy fibers. These results suggest that parvalbumin appears to have more protective effect against neuronal loss and that mossy fiber synaptic reorganization seems to play a major role in pathogenesis of hippocampal sclerosis of human temporal lobe epilepsy.

Evaluation of Prognostic Significance of AgNORs and PCNA during 9,10-dimethyl-1,2-benzantracene(DMBA)-induced Hamster Buccal Pouch Carcinogenesis.: Sam Pyo Hong, Myong Soon Song, Seong Doo Hong, Jae Il Lee, Chang Yun Lim; Korean J Pathol. 1998;32(5):337-345.

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Abstract: The purpose of this study is to evaluate the prognostic significance of argyrophilic nucleoalr organizer regions (AgNORs) and proliferating cell nuclear antigen (PCNA) by using DMBA hamster buccal pouch carcinogenesis which provides a good experimental model in reproducing steps from precancerous lesions to invasive squamous cell carcinomas. The buccal pouches of 50 Syrian hamsters were applied with 0.5% DMBA in mineral oil three times a week to reproduce various lesions from precancerous ones such as hyperkeratosis or epithelial dysplasia to invasive squamous cell carcinomas. Their sections were stained with H & E, and silver colloid, and processed immunohistochemically by being applied with monoclonal antibody to PCNA. The histopathologic examainations were done and the counts of AgNORs were evaluated. The PCNA labelling indices on each lesions were evaluated. The correlation between histopathological grades and counts of AgNORs or PCNA labelling indices were evaluated. The number of AgNORs was 2.22+/-0.22 in control group, 3.46+/-0.72 in carcinoma in situ (CIS), 3.78+/-0.63 in squamous cell carcinoma (SCC), respectively. AgNORs significantly increased in severe epithelial dysplasia, CIS, and SCC compared with normal tissue (P<0.05). The PCNA Labeling Index (LI) was 39.47+/-6.68% in control group, 79.61+/-4.14% in CIS, and 85.43+/-6.25% in SCC, respectively. PCNA LI also significantly increased in epithelial dysplasia, CIS, and SCC compared with normal tissue (P<0.05). The number of AgNORs, AgNOR area, and PCNA LI slightly increased in the advancing front than in the center of SCC, but, it was not statistically significant. It appeared that there were a good correlation between the number of AgNORs and PCNA LI (Pierson correlation coefficient : 0.649, P<0.001). These results suggested that the number of AgNORs and the PCNA LI could be useful markers for evaluating the risk of malignant transformation and prognosis of SCC. It was thought that the clinical usefulness of these markers should be verified by using human tissue specimens.

Estrogen and Progesterone Receptor Expressions in Benign Prostatic Hypertrophy and Prostatic Adenocarcinoma.: Mi Seon Kang, Seo Young Park, Hye Kyoung Yoon; Korean J Pathol. 1998;32(5):346-351.

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Abstract: The effect of androgen in the development of the normal prostate and the evolution of benign prostatic hypertrophy (BPH), and prostatic adenocarcinoma has been proven. In addition to androgen, estrogen and progesterone are also thought to play a role in the pathogenesis of BPH and carcinoma. However, their exact roles are not yet known because there is no conclusive evidence. Thirty cases of prostatic adenocarcinoma and 16 cases of BPH were studied. Immunohistochemical staining for estrogen receptor (ER) and progesterone receptor (PR) in epithelial and stromal cells, respectively was performed and the results were assessed semiquantitatively based on the number of positive cells per 100 total cells. Slides were scored as negative; less than 5% of cells, 1 ; 6~15% of cells, 2 ; 16~25% of cells, and 3 ; more than 26% of cells. The relationship between ER and PR expression and the patient's age, histologic grade, and clinical stage was evaluated in prostatic adenocarcinomas. ER was negative in epithelial and in stromal cells for all prostatic adenocarcinomas and BPH cases. The PR expression in epithelial cells and in stromal cells of BPH was noted in 15 (93.8%) and 16 (100.0%) out of 16, respectively. The PR expression of carcinoma cells and stromal cells in prostatic adenocarcinoma was found in 28 (93.3%) and 23 out of 30 (76.7%), respectively. The PR immunoreactivities of stromal cells around carcinoma were 3 in 18 cases, 2 in one case, and 1 in 4 cases, but those of epithelial and stromal cells of BPH and carcinoma cells of prostatic carcinoma were similar to each other with a value of 3 in most cases. The PR expression rate of stromal cells around carcinoma was significantly correlated with the patient's age (p=0.044), but not with histologic grade and clinical stage. In summary, estrogen does not have a direct effect on the biological behavior of BPH and prostatic adenocarcinoma, but progesterone appears to play a role in the pathogenesis of BPH and prostatic adenocarcinoma. Further studies should clarify the biological role of progesterone in the human prostate.

A Study for IL-6, IL-13 and TIMP-3 Expressions of Placenta, Fetus and Endometrium in Pregnant Mice after Treatment with Lipopolysaccharide.: Sung Ran Hong, In Gul Moon, Ju Young Seoh, Yee Jeong Kim, Sung Sook Kim, Woon Sup Han; Korean J Pathol. 1998;32(5):352-361.

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Abstract: We examined C3H pregnant mice at 15 days (70% gestation) after treatment of lipopolysaccaride (LPS) to observe the changes of IL-6 concentration in maternal serum and amniotic fluid and expression of IL-6, IL-13 & TIMP-3 in placenta, fetus and endometrium, and to investigate the correlation among IL-6, IL-13 and TIMP-3. The results were as follows: 1) IL-6 in serum and amniotic fluid after treatment of LPS was significantly elevated; peaked at 1, 2, 4, 5 hours and decreased to control level at 8 hours (P<0.05). IL-6 in placental disc, chorioamnionic membrane, fetus, decidua and endometrial epithelium was overexpressed significantly at 1, 2, 4 hours after treatment of LPS (P<0.05). IL-6 overexpression was more significantly increased in maternal tissue than fetal tissue (P<0.05). 2) Increased concentration of amniotic fluid IL-6 was equally originated from transplacental crossage of maternal serum IL-6, and direct local production of IL-6 from placenta, fetus and endometrium (P<0.05). 3) IL-13 in placental disc, chorioamnionic membrane, fetus, decidua and endometrial epithelium was overexpressed after treatment of LPS, but not significant statistically. 4) TIMP-3 was overexpressed in placental disc, chorioamnionic membrane, fetus and decidua. TIMP-3 overexpression was more significant in placental disc than other tissues (P<0.05). 5) Overexpressions in IL-13 and IL-6 revealed direct proportional correlation coefficient (Spearman correlation coefficient, 0.5212 ; P<0.05). IL-6 expression was a head of overexpression of TIMP-3, but not significant. In conclusion, all of IL-6, IL-13 and TIMP-3 relate with inflammatory response, especially IL-6 in maternal serum, amniotic fluid and tissue of placenta, fetus and endometrium was so sensitive that it can be an indicator for antenatal diagnosis of chorioamnonitis, and amniotic fluid IL-6 is equally originated from maternal serum and from tissue of placenta, fetus and endometrium. IL-13 and TIMP-3 may have parallel correlation to the IL-6 in fetal and maternal tissue after treatment of LPS.

Carcinosarcoma (Malignant M llerian Mixed Tumor) of the Female Genital Tract: A clinical and pathologic study of ten carcinosarcomas.: Sung Ran Hong, Yee Jeong Kim, Hy Sook Kim, Jae Uk Shim, Chong Taik Park; Korean J Pathol. 1998;32(5):362-369.

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Abstract: Carcinosarcomas of the female genital tract have generally been regarded as a type of sarcoma. Recent studies, however, suggest the tumor may be more closely related to carcinoma and may represent metaplastic carcinoma in histogenesis. We analyzed clinicopathologic and immunohistochemical features of 10 carcinosarcomas to evaluate the relative importance of the carcinomatous and sarcomatous components in metastasis and recurrence. The primary tumor originated in the uterine body in seven cases, the uterine cervix in two and the ovary in one. Patient,s ages ranged from 54 to 71 years (mean, 64). The most common symptom of the uterine mass was vaginal bleeding. The median survival time was 21 months following diagnosis in five cases. Surgico-pathologic FIGO stages of five patients who received an operation were stage III and IV, but clinical FIGO stage of three patients (60%) among them were I. Lymphovascular invasions were identified in seven areas; five vascular invasion lesions showed the carcinomatous component alone, one the sarcomatous component alone, and remained one admixture of both components. Metastatic and recurrent lesions to the paraaortic lymph node, ovary, pelvic wall, or vaginal vault showed characteristically carcinomatous component only. Immunohistochemically, positive reactions for cytokeratin and epithelial membrane antigen were noted in the sarcomatous component of five cases. Vimentin positivity was detected in carcinomatous component of three cases. We conclude that the dominant element in carcinosarcomas of the female genital tract is the carcinomatous component. The survival rate of carcinosarcoma is extremely poor. The surgico-pathologic stage is better indicator of survival than the clinical stage. Immunohistochemical findings suggest that carcinosarcoma may represent a metaplastic carcinoma in histogenesis.

Carcinosarcoma of the Female Genital Tract: Immunohistochemical study on transitional area further supports the metaplastic origin.: Chan Pil Park, Joo Seob Keum, Gu Kong, Moon Hyang Park, Jung Dal Lee; Korean J Pathol. 1998;32(5):370-377.

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Abstract: Carcinosarcoma of the female genital tract, also called malignant mixed mullerian tumor (MMMT), is a rare and relatively aggressive tumor with various homologous and heterologous components. There have been various studies to find prognostic factors and to investigate the histogenesis, including tissue culture, electron microscopy, and immunohistochemical studies. We investigated carcinomatous, sarcomatous, and transitional areas of 6 cases of carcinosarcoma of the uterus and ovary by using epithelial and mesenchymal markers. Immunohistochemical profiles of the transitional areas were significantly different from those of carcinomatous and sarcomatous areas. Immunoreactivities for cytokeratin and epithelial membrane antigen (50% and 22.2%) were weaker than those of carcinomatous areas (95.2% and 100%), but stronger than those of sarcomatous areas (11.1% and 5.6%)(p<0.01). In transitional areas, vimentin, smooth muscle actin and S-100 protein were more strongly expressed than in carcinomatous areas, but more weakly expressed than in sarcomatous areas (p<0.01, p<0.01, and p=0.018, respectively). Myoglobin was entirely negative in carcinomatous areas and immunoreactive in minor portions of transitional and sarcomatous areas (22.2% and 16.7%, respectively). These results suggest that the transitional areas are between the carcinomatous and sarcomatous nature in differentiation, further supporting that the carcinosarcomas of the female genital tract may arise, through metaplastic change, from a type of carcinoma.

Sarcomatoid Renal Cell Carcinoma; Special Reference to its Distinction from Carcinosarcoma.: Kee Taek Jang, Yeon Mee Kim, Je Geun Chi; Korean J Pathol. 1998;32(5):378-381.

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Abstract: Sarcomatoid renal cell carcinoma is an uncommon tumor that has to be distinguished from renal carcinosarcoma. We have described three cases of sarcomatoid renal cell carcinoma showing different clinical and light microscopic features. An ultrastructural study of the tumor cells from the sarcomatoid area revealed frequent desmosomal junction, confirming the epithelial nature of the neoplasm. All three cases showed an aggressive clinical course and tended to invade adjacent organs or tissues. We believe that an histological and immunohistochemical examination in conjunction with an electron microscopic examination are necessary to diagnose sarcomatoid renal cell carcinoma.

Case Reports

Pulmonary Endometriosis: A case report.: Soo Jin Jung, Young Ju Kim, Hye Kyoung Yoon; Korean J Pathol. 1998;32(5):382-384.

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Abstract: Pulmonary endometriosis is a rare disease which is characterized by hemoptysis during menstruation (catamenial hemoptysis). We report a case of pulmonary endometriosis in a 33-year-old housewife. She has had regular menses with moderate flow and minimal dysmenorrhea. She had undergone curettage in May 1995 for artificial abortion. In July 1995, she experienced the first episode of hemoptysis. A chest CT scan revealed a 2.0 1.0 cm sized ill-defined soft tissue density in the periphery of anterior segment of the left upper lobe with a surrounding irregular ground-glass opacity. A left upper lobectomy was done under the diagnosis of pulmonary endometriosis. Cut section of the resected lung showed a round red-brownish solid lesion, measuring 2.0x1.0cm in cross. Microscopically a focus of the endometrial tissue, which was composed of endometrial glands and stroma, was found in the lung parenchyme and many hemosiderin laden macrophages were seen in the surrounding alveoli. The postoperative course was favorable with no further episodes of hemoptysis.

Adreno-Hepatic Fusion: A case report.: Kyung Moo Yang, Young Nyun Park, Chan Il Park; Korean J Pathol. 1998;32(5):385-387.

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Abstract: Adreno-hepatic fusion is rare condition defined as adhesion of the liver and right adrenal cortex with close intermingling of the respective parenchyme. It is suggested to be an aging phenomenon, because its incidence is much higher in older age group. Clinically it may pose a problem of operability of the organ involved. We report a case of incidentally found adreno-hepatic fusion in a 49 year old female patient with adenocarcinoma of the sigmoid colon. The segementectomy of VIII segement of the liver was done due to a 6 4 cm sized metastatic nodule of adenocarcioma. Pathological examination of the liver revealed an ovoid shaped, 1 0.5 cm sized adrenal cortical tissue. It was subcapsularly located and about 1cm apart from the metastatic adenocarcinoma with an intervening normal hepatic tissue. The adrenal tissue was mainly composed of zona fasciculata without medullary tissue. In the interphase, the adrenal tissue and liver tissue were admixed closely and partially septated by thin fibrous tissue. There was no inflammatory response to the heterotropically located adrenal tissue and there was no symptom related to the adrenal gland.

Polypoid Ganglioneuromatosis of Colon: A case report.: Jin Ja Park, Kyung Chan Choi, Young Hee Choi, Young Euy Park; Korean J Pathol. 1998;32(5):388-390.

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Abstract: Gastrointestinal ganglioneuromatosis is an extremely rare lesion which typically occurs with a significant systemic syndrome. It is known to be a major component of multiple endocrine neoplasia, type 2b. We presented a case of polypoid ganglioneuromatosis of the colon in a 3-year-old female with abdominal pain and diarrhea. She had no clinical evidence of the systemic syndrome or von Recklinghausen's neurofibromatosis, conditions in which intestinal ganglioneuromatosis can occur. Gross examination showed diffuse polypoid masses in ascending and transverse colons with normal-appearing mucosa. Microscopic examination revealed a proliferation of spindle-shaped neuronal cells containing multiple clusters of mature ganglion cells in the mucosa, submucosa and proper muscle. We describe a case of colonic ganglioneuromatosis without any component of multiple endocrine neoplasia or family history.

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