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1.
Chinese Journal of Oncology ; (12): 118-120, 2009.
Article in Chinese | WPRIM | ID: wpr-255549

ABSTRACT

<p><b>OBJECTIVE</b>To detect the expession of THY1 in ovarian serous cystadenocarcinoma tissues.</p><p><b>METHODS</b>Immunohistochemistry was performed to detect the expression of THY1 gene in formalin-fixed, paraffin-embedded specimens of normal ovaries (n = 25), ovarian serous cystadenoma (n = 25), and serous cystadenocarcinoma (n = 53). The correlation of THY1 expression with clinicopathological parameters was statistically analyzed.</p><p><b>RESULTS</b>The positive expression rates of THY1 protein in normal ovaries, ovarian serous cystadenomas and ovarian serous cystadenocarcinomas were 60.0% (15/25), 72.0% (18/25) and 34.0% (18/53), respectively. The values of IOD of THY1 protein expression were 288,449.2 +/- 60,087.3, 271,655.6 +/- 66,588.7 and 252,087.6 +/- 45,559.4, respectively. The expression of THY1 protein was significantly down-regulated in ovarian serous cystadenocarcinoma tissues compared with that in normal ovarian tissues and ovarian serous cystadenoma tissues (P < 0.05). THY1 expression was negatively correlated with surgical-pathological staging, histological differentiation and lymph node involvement (P < 0.05).</p><p><b>CONCLUSION</b>The decreased level of THY1 expression may be related with the occurrence and development of ovarian serous cystadenocarcinoma.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Cystadenocarcinoma, Serous , Metabolism , Pathology , Cystadenoma, Serous , Metabolism , Pathology , Gene Expression Regulation, Neoplastic , Immunohistochemistry , Lymphatic Metastasis , Neoplasm Staging , Ovarian Neoplasms , Metabolism , Pathology , Thy-1 Antigens , Metabolism
2.
Journal of Southern Medical University ; (12): 84-91, 2007.
Article in Chinese | WPRIM | ID: wpr-298235

ABSTRACT

<p><b>OBJECTIVE</b>To construct THY1 eukaryotic expression plasmid and study its effects on the growth of epithelial ovarian cancer cell line SKOV3.</p><p><b>METHODS</b>THY1 gene fragment was obtained from normal human ovarian tissue using RT-PCR, and inserted into the eukaryotic expression plasmid pcDNA3.1(+) to construct the recombinant plasmid pcDNA3.1(+)-THY1, which was transformed into E. coli JM109 followed by selection of the positive clones containing the target inserts. The eukaryotic expression plasmid was analyzed by PCR, restriction endonucleases digestion and DNA sequencing. SKOV3 cells divided into SKOV3-THY1, SKOV-3-Null and SKOV3 groups were transfected via liposome with the recombinant plasmid pcDNA3.1(+)-THY1, empty plasmid, or not transfected, respectively. The expression of THY1 mRNA and its protein were examined by RT-PCR and Western blot methods. The cell growth and apoptosis were evaluated by MTT assay and flow cytometry.</p><p><b>RESULTS</b>The gene fragment of exogenous THY1 was correctly inserted into the eukaryotic expression plasmid pcDNA3.1(+) as verified by PCR, restriction endonucleases digestion and DNA sequencing, and recombinant expression plasmid pcDNA3.1(+)-THY1 transfection resulted in stable expression in SKOV3 cells as shown by RT-PCR and Western blotting. The cell growth inhibition rate of SKOV3-THY1 group (56.6% at the fifth day) was significantly higher than that of the SKOV3-Null group (12.5%, P<0.05), and the cell apoptosis rate in SKOV3-THY1 group (31.8%) was significantly higher than those in SKOV3-Null group (10.5%) and SKOV3 group (9.8%, P<0.05), but the apoptosis rate between the latter two groups was similar (P>0.05).</p><p><b>CONCLUSIONS</b>The recombinant plasmid pcDNA3.1(+)-THY1 can be expressed stably in human ovarian cancer cell line SKOV3. THY1 transfection can inhibit the growth of SKOV3 cells in vitro, suggesting the important role of THY1 gene in pathogenesis and development of ovarian cancer.</p>


Subject(s)
Female , Humans , Apoptosis , Genetics , Physiology , Blotting, Western , Cell Line, Tumor , Cell Proliferation , Cell Survival , Genetics , Physiology , Epithelial Cells , Metabolism , Pathology , Flow Cytometry , Gene Expression , Ovarian Neoplasms , Genetics , Metabolism , Pathology , Plasmids , Genetics , RNA, Messenger , Genetics , Metabolism , Recombinant Proteins , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Thy-1 Antigens , Genetics , Physiology , Transfection
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