| ObjectiveThe ovarian malignant tumor is one of the commonest malignant tumors in female reproductive organs. Its incidence is the third in malignant tumor of female reproductive organs, and lowers than that of cervical cancer and endometrial cancer. Because the anatomy position of the ovary is deep, its tumor is difficult to be found early and it is poor to treat . Although cytoredutive operation of the ovarian cancer is improved greatly and new chemical therapeutic drugs are developed and used, the 5 - years survival of ovarian carcinoma is very low, a-bout 25% -30%. Therefore, there is increasing urgency to develop new and effective strategies to prevent and treat ovarian carcinoma.Nonsteroid and - inflammatory drugs ( NSAIDs) were first found by Felix Hoffman in 1893 and used as the treatment of arthritis. For a long time, NSAIDs have been used as analgesic and anti - inflammatory drugs in clinical. Recently they were found to be able to prevent colon cancer. A large number of researches indicate that NSAIDs can inhibit the proliferation of tumor cells, and promote apoptosis of tumor cells. Therefore NSAIDs is believed to have potential value in chemical prevention. Vane reported in 1971 that the anti - inflammatory mechanisms of NSAIDs were inhibition of cyclooxygense ( COX) , which is the key enzyme in the conversion of arachidonic acid ( AA) to prostaglandins. Two isoforms of COX, namely COX - 1 and COX -2, have been identified, each of which is encoded by individual gene, located on different chromosomes. According to the different inhibitioneffects on COX, there are three kinds of NSAIDs, 1 ) selective COX - 1 inhibitors, 2) selective COX - 2 inhibitors, 3 ) non - selective COX inhibitors. NS398, a derivative of sulfamilamide, is belong to selective COX -2 inhibitors and can specially inhibit COX -2.Recent studies have reported that the expression of COX - 2 is up regulation in colon cancer, pancreatic cancer, lung cancer, prostate cancer, gastric cancer and so on, while the expression of COX ?1 is unaltered. We also observed that there was expression of COX - 2 in ovarian epithelial serous carcinoma, in which the expression rate was 81. 9% . To investigate if NS398 can inhibit human ovarian carcinoma cells and provide experimental data to the prevention and treatment of ovarian cancer, we observed the effect of NS398 on human ovarian cacinoma cell lines in vitro.Materials and MethodsMaterials. Two ovarian epithelial serous carcinoma cell lines, CAOV3 and OVCAR3, were provided by cytobiological lab of China Medical University. CAOV3 and OVCAR3 were cultured respectively in RPMI1640 containing 10% FBS, 100u/ml penicillin and 100ug/ml streptomycin, at 37C ,5% CO2 in a constant - temperature enclosure CO2 incubator. Then digested by 0. 25% Trypsin.Reagents. NS398 was purchased from American Cayman chemical company. Anti - COX -2 monoclone antibody and SP immunohis-tochemical kit were purchased from American Santa Cruz Inc. The primers of COX - 2 mRNA and b - actin mRNA were synthesized by Beijing Aoke biological Inc. Reverse transcription kit was purchased from TaKaRa Inc. MTT and trypan blue were purchased from Ameri-can Sigma Inc. TdT apoptosis in situ staining kit was purchased from Wuhan Boside Biological Inc. RT - PCR and DNA ladder reagents were provided by the congenital malformation lab of the second clinical college of China Medical University. Methods.1. Immunohistochemical assay.2. RT-PCR assay.3. Experiment of cell growth inhibition. 3. 1 CeU number assay3. 2 MTT assay.4. Cell morphology assay.4. 1 Inverted phase contrast microscope.4. 2 Electron microscope.5. DNA - ladder electrophoresis.6. Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL).Statistical Methods.Statistical analysis was performed using SPSS for windows 10. 0 software. The results of human ovarian carcinoma cell lines treated with NS398 was expressed as mean ?s. And comparisons of means were carried out by one w... |
PDF Full Text Request