CATHEPSIN B EXPRESSION AND ITS RELATIONSHIP WITH MICROVESSEL DENSITY AND BIOLOGICAL BEHAVIOUR OF COLORECTAL CARCINOMA

Objective: To investigate cathepsin B(CB) expression in colorectal carcinoma and its relationship with microvessel density (MVD) and biological behavior. Methods: CB and MVD were detected by immunohistochemistry in 47 cases of colorectal carcinoma. Results: The expression of CB in mucinous colorectal carcinoma was significantly higher than that in no-mucinous colorectal carcinoma. There was significant difference (P<0.05). The MVD in group with positive CB was stronger than that in group with negative CB. There was also significant difference (P<0.05). Conclusion: The results suggest that CB expression has correlation with MVD, invasion and metastasis in colorectal carcinoma, especially in mucinous colorectal carcinoma.

Expression of Wnt-1,beta-catenin and c-myc in Ovarian Epithelial Tumor and Its Implication

Objective： To investigate the expression of Wnt-1, beta-catenin and c-myc in normal ovarian epithelial cell and malignant ovarian epithelial tumor. Methods： Immunohistochemical staining with SP method was conducted to identify the expression of Wnt-1, beta-catenin and c-myc in 18 samples of normal epithelial tissue and 34 cases of malignant epithelial tumor of ovary. Results： The expression rate of Wnt-1 and c-myc in malignant epithelial tumors was higher than those in normal epithelial cell （P〈0.05）. The abnormal expression rate of beta-catenin in malignant ovarian epithelial tumors was higher than that in normal epithelial cell （P〈0.05）. A significant positive correlation was found between Wnt-1, beta-catenin and c-myc in malignant ovarian epithelial tumor （P〈0.05）. A significant difference of expressions of Beta-catenin and C-myc was found between serous and mucinous tumors （P〈0.05）. Conclusion： The abnormal expression of Wnt-1, beta-catenin and c-myc might indicate the malignant transformation in ovarian epithelial tumors.

Clinicopathological Features of Extraskeletal Myxoid Chondrosarcoma:An Analysis of 9 Cases

Objective： To investigate the Clinicopathological （EMC）. Methods： Nine cases of extraskeletal characteristics of extraskeletal myxoid chondrosarcoma myxoid chondrosarcoma were studied. Extensive immunohistochemical analysis was performed in all the cases and ultrastructural studies were done in 2 extraskeletal myxoid chondrosarcomas. Follow-up information was available for seven patients. Results： There were 7 males and 2 females whose ages ranged from 31 to 69 years （median 52.78 years）. Local pain or tenderness and the presence of a palpable mass were the main complaints of the patients. The tumors were located mainly in the lower extremities （66.7%）. Most tumors were deep-seated. They usually had a distinct multinodular configuration delineated by fibrous connective tissue. The tumor cells were arranged in delicate intersecting strands, rings, and garlands for the most part. The myxoid matrix was abundant in most cases. Immunohistochemical analysis was performed in all the cases and ultrastructural studies were done in 2 extraskeletal myxoid chondrosarcomas. EMC expressed vimentin （100%, 9/9）, neuron-specific enolase （77.8%, 7/9）, S-100 protein （66.7%, 6/9）, synaptophysin and chromogranin A （22.2%, 2/9）. None of the tumors expressed EMA and desmin. Ultrastructurally： EMC was characterized by distinct cords of cells immersed in a glycosaminoglycan rich matrix. The cells were rich in mitochondria, had well-developed Golgi apparatus and there were numerous smooth vesicles. In many cells, there were also prominent glycogen deposits and lipid droplets. Some tumor cells had intracisternal microtubules. In one of the 2 extraskeletal myxoid chondrosarcomas there were 140-180 nm diameter membrane-bound dense-core secretory granules in cell bodies. Conclusion： Extraskeletal myxoid chondrosarcoma （EMC） is a rare soft tissue sarcoma characterized by distinctive morphological and cytogenetical features. However, the chondroid nature has been a subject of controversy, and its line of differentiation remains to be determined. A substantial proportion of EMC shows immunophenotypic and/or ultrastructural evidence of neuroendocrine differentiation. EMC has high potential of local recurrence and metastasis, and a high disease-associated death rate.

Expressions of beta-catenin, APC Protein, C-myc and Cyclin D1 in Ovarian Epithelial Tumor and Their Implication

Objective： To investigate the expressions of beta-catenin, protein APC （adenomatous polyposis coil protein）, c-myc and cyclin D1 and their implication in ovarian epithelial tumor. Methods： Immunohistochemical staining with SP method was conducted to identify the expressions of beta-catenin, APC protein, c-myc and cyclin D1 in ovarian epithelial tumor in 48 cases. Results： The abnormal expression rate of beta-catenin in malignant and borderline ovarian epithelial tumors was higher than that in benign epithelial tumors （P〈0.01）. The expression rates of c-myc and cyclin-D1 in ovarian malignant and borderline epithelial tumors were higher than those in benign epithelial tumors too（P〈0.05）. The prevalence of APC protein positive expression in benign epithelial tumors were significantly greater than that in malignant epithelial tumors （P〈0.05）. A significant negative correlation was found between beta-catenin and APC protein in ovarian epithelial tumors; while a significant positive correlation was found between beta-catenin, c-myc and cyclin-D1 in ovarian epithelial tumor （P〈0.05）. Conclusion： The abnormal expressions of Beta-catenin, APC protein, c-myc and cyclin-D1 might be used to indicate the malignance transform of ovarian epithelial tumors.

Expressions of Beta-catenin,E-cadherin and MMP-7 in Ovarian Epithelial Tumors

Objective： To investigate the expressions of Beta-catenin, E-cadherin and MMP-7 and their implications in ovarian epithelial tumor. Methods： Immunohistochemical staining with SP method was conducted to identify the expressions of Beta-catenin, E-cadherin and MMP-7 in ovarian epithelial tumor in 66 cases. Results： The abnormal expression rate of Beta-catenin in malignant ovarian epithelial tumor was higher than those in borderline and benign epithelial tumors （P〈0.05）. The positive rates of E-cadherin in benign and borderline ovarian epithelial tumors were significantly greater than that in malignant epithelial tumor. The expression rates of MMP-7 in malignant and borderline ovarian epithelial tumors were higher than that in benign epithelial tumor （P〈0.05）. Conclusion： The abnormal expressions of Beta-catenin, E-cadherin and MMP-7 might be used to indicate the malignance transform of ovarian epithelial tumors, but they have no significant correlation with peritoneal dropsy invasion, caul invasion and appendant invasion in ovarian epithelial tumor.

Relationship between Expression of beta-catenin and VEGFs(VEGFA,VEGF-C),VEGF Receptors-2(VEGFR-2)in Medulloblastoma

Objective： To investigate the expression of beta-catenin and VEGFs （VEGF-A, VEGF-C） and VEGF receptor-2 （VEGFR-2） protein in medulloblastoma. Methods： Immunohistochemical staining with SP method was conducted to determine the expression of beta-catenin and VEGFs （VEGF-A, VEGF-C） and VEGFR-2 in 33 cases of medulloblastoma and 10 normal cerebellar tissues. Results： The expression rate of beta-catenin, and VEGFs （VEGF-A, VEGF-C） and VEGFR-2 in medulloblastoma were significantly higher than that in normal tissue. A significant positive correlation was found between beta-catenin and VEGFs （VEGF-A, VEGF-C） and VEGFR-2 protein in medulloblastoma. Conclusion： There was a correlation between beta-catenin and VEGFs （VEGF-A, VEGF-C） and VEGFR-2 in medulloblastoma, which may play a role in the pathogenesis and development of medulloblastoma.

Expression of Beta-Catenin and APC Protein in Ovarian Epithelial Tumor and Its Implication

Objective： To investigate the expression of beta-catenin, APC protein and its implication in ovarian epithelial tumor. Methods： Immunohistochemical staining with SP method was conducted to determine the expression of beta-catenin and APC protein in 48 cases of ovarian epithelial tumor. Results： The abnormal expression rates of beta-catenin in ovarian malignant and borderline epithelial tumors were higher than that in benign epithelial tumors. The expression of APC protein in benign epithelial tumors was significantly greater than that in malignant epithelial tumors. A significant negative correlation was found between beta-catenin and APC protein in ovarian epithelial tumors. Conclusion： Beta-catenin and APC protein have important effect on pathogenesis and development of ovarian epithelial tumors.

Expression of Maspin,uPA and MMP-7 in Human Gastric Carcinoma

Objective： To investigate the expression of maspin, uPA and MMP-7 in gastric carcinoma and discuss their roles in tumorigenesis and progression of gastric cancer. Methods： lmmunohistochemistry SP method was used to detect expression of maspin, uPA and MMP-7 in gastric adenocarcinoma （n=30）, signet-ring cell carcinoma （n=30）, normal gastric mucosa （n=20）; in addition, reverse transcriptase PCR was carried out to compare the mRNA expression of maspin between gastric adenocarcinoma and normal mucosa. Results： The expression rates of maspin, uPA and MMP-7 protein were 50%, 70% and 80% in gastric adenocarcinoma, and 46.7%, 76.7%, and 90% in signet-ring cell carcinoma, respectively. However, the expression rates of maspin, uPA, and MMP-7 protein were 90%, 35% and 30%, respectively in normal gastric mucosa. The relative transcription level of maspin mRNA in normal gastric mucosa was higher than that in gastric adenocarcinoma （P〈0.01）. Maspin protein expression was significantly related to the depth of invasion and lymph node metastasis of gastric carcinoma, but did not relate to tumor size and TNM staging. The protein expression of uPA and MMP-7 were significantly related to invasive depth, lymph node metastasis and TNM staging of carcinoma, but not to tumor size. Maspin protein expression had a negative correlation with uPA and MMP-7, while uPA protein expression had a positive correlation with MMP-7. The relative mRNA transcription level of Maspin significantly correlated with the lymph node metastasis and the depth of invasion of carcinoma, but did not relate to TNM staging or tumor size. Conclusion： Down-regulated expression of maspin and up- regulated expression of uPA and MMP-7 play important roles in the invasion and metastasis of gastric carcinoma. They may serve as effective markers of the biopathological behavior of gastric tumor.

THE FURTHER STUDY ON BIOLOGICAL BEHAVIOR OF COLORECTAL ADENOCARCINOMA:THE EXTRACELLULAR PROTEOGLYCAN DEGRADED BY ARYLSULFATASE B

The fresh tissues were obtained from 64 colorectal adenocarcinoma (43 well-differentiated and moderately-differentiated adenocarcinoma, 12 poorly- differentiated adenocarcinoma and 9 mutinous cell carcinoma including signet ring cell carcinoma) during surgical operation. The resected edge of each specimen was used for control group. The arylsulfatase B was studied by histochentical staining in different types of colorectal adenocarcinoma, among which 19 cases were investigated by electronhistochemical staining so as to observe the Ruthenium Red granules alteration which represented the extracellular proteoglycan changes and ultrastructure of cancer cells.The results showed that the mutinous cell carcinoma was of the most Intensive arylsulfatase B activity and has a lot of secretory granules with various electron densities in the cytoplasm. The Ruthenium Red granules close to the cancer cell disappeared, a part of remainders changed into the lowered electron density and indistinct shape. In contrast, the other types adenocarcinoma revealed less enzyme activity and a fewer secretory granules. The Ruthenium Red granules near the cancer nest showed that their electron density and size were identical with those of the control group. All of these mentioned above indicate that mucinouscell carcinoma may release hydrolase into pericancerous matrix to degrade the proteoglycans. In view of the network structure formed by proteoglycan in the connective tissue, network has ability to hinder the cancer cell spreading. Because the arylsulfatase B is able to degrade the dermatan sulfate proteoglycan which is component of proteoglycans in the extracellular matrix of human colon. We consider that the arylsulfatase B may lead to destruction of the network barriers in the connective tissue in favour of cancer cell invasion. So the mucinous cell carcinoma is more malignant than those of other colorectal adenocarcinoma.

EXPERIMENTAL STUDY OF THE DIFFERENTIATION HPV16 SUBGENES-IMMORTALIZED HUMAN ENDOCERVICAL CELLS INDUCED BY ALL-TRANS-RETINOIC ACID

Objective： To investigate the differentiation-inducing effects of all-trans-retinoic （ATRA） to HPV16 subgenesimmortalized human l：ndocervical cells （H8 cell） in vitro. Methods： HPV16 subgenes-immortalized human endocervical cells （H8 cells） were cultured in vitro. After treated with ATRA, the proliferation of immortalized human endocervical cells was measured by MTT assay; morphological changes were observed using M and TEM; cell cycle was analyzed by FCM; expression of Ki67 was tested using immunocytochemistry and the activity of telomerase was tested using PCR-ELISA. Results： ATRA could inhibit proliferation of H8 cells significantly and induce their morphodifferentiation. According to FCM, H8 cells accumulated in G1 phase and expression of Ki67 and activity of telomerase reduced significantly after treatment with ATRA. Conclusion： ATRA could induce the differentiation of H8 cell line obviously, which might be achieved by inhibiting proliferation, blocking cell cycle, and reducing activity of telomerase.

Inhibitory Effect of Nordy on HPV16 E6 Gene in Human Immortalized Endocervical Cells

Objective： To study the inhibitory effect of Nordy on HPV16 E6 gene in human immortalized endocervical cells （H8）. Methods： The expressions of E6 and p53 proteins of H8 cells were detected by immunocytochemistry, and the expression of HPV16E6 mRNA of H8 cells was detected by RT-PCR. Results： According to immunocytochemistry, expressions of E6 and p53 proteins of H8 cells after treatment with 100 μtmol/L Nordy for 72 h were weaker than control group obviously. After treatment with 50, 100, 200 μmol/L Nordy for 72 h. HPV16E6 mRNA expressions of H8 cells were lower than the control group significantly （P〈0.05）. Conclusion： Nordy could inhibit the expression of HPV16 E6 mRNA in H8 cells, influence transcription of E6 gene and result in decreased expressions of E6 and p53 proteins.

CHANGES OF NUCLEAR MATRIX PROTEIN AND ITS RELATIONSHIP WITH c-erbB-2 IN HUMAN COLON ADENOCARCINOMA

Objective： Nuclear matrix protein is tissue, cell-type specific, and tumor-relative. It plays an important role in the regulation of intranuclear processes. Some researches also showed that a c-erbB-2 promoter-specific DNA-binding nuclear matrix protein is present only in malignant human breast tissues and induces mitogenesis and cell surface expression of the c-erbB-2 protein in resting NIH/3T3 cells. But it is not clear that how it in colon adenocarcinomas. Methods： Two-dimensional gel electrophoretic method was used for NMP identification and immunohistochemistry was used for c-erbB-2 detection in 12 cases of colon adenocarcinomas and matched adjacent normal colon tissues. Results： 5 different nuclear matrix proteins （named C1-C5） were identified in 12 colon adenocarcinoma specimens, but not in the matched adjacent normal colon tissues; 3 nuclear matrix proteins （named N1-N3） were identified in all 12 matched adjacent normal colon tissues, but not in colon adenocarcinoma specimens. A nuclear matrix protein （named N4） was detected in all of 9 moderated-well differentiated adenocarcinomas and all 12 matched adjacent normal colon tissues, but not in 3 poor-differentiated adenocarcinomas. All of the 10 colon adenocarcinomas which had the nuclear matrix protein C4 were c-erbB-2 expression positive. Conclusion： The data suggest that there are specific nuclear matrix proteins in colon adenocarcinomas and its subtypes, which maybe valuable to serve as markers of colon adenocarcinomas in future. Nuclear matrix protein C4 probably is a c-erbB-2 promotor-specific nuclear matrix protein in colon adenocarcinomas, and may induce the expression of c-erbB-2.

Effects of Nordy on the proliferation, differentiation,and apoptosis of HPV16 subgene-immortalized human endocervical cells

Objective The aim of this study was to investigate the effects of Nordy on the proliferation, differentiation, and apoptosis of HPV16 subgene-immortalized human endocervical cells(H8 cells).Methods After treatment with Nordy, H8 cell proliferation was evaluated using the MTT assay. The effects of Nordy on the cell cycle and apoptosis of H8 cells were analyzed by flow cytometry(FCM) and the Annexin V-FITC method. H8 cell MCM5 expression was detected by immunocytochemistry. Morphological changes were observed by light and electron microscopy. Telomerase activity was evaluated by TRAP-ELISA.Results We found that 10 μmol/L–100 μmol/L Nordy significantly inhibited H8 cell proliferation. After treatment with Nordy, H8 cells were blocked in the G0/G1 phase, and the rate of cell apoptosis increased significantly. Cells differentiated toward innocuousness, and MCM5 expression and telomerase activity notably decreased.Conclusion Nordy was observed to inhibit proliferation and promote apoptosis in H8 cells. Nordy also induced H8 cell differentiation; this effect may have been achieved by blocking the cell cycle and decreasing telomerase activity.