On Effects Imposed By NS-398 On Expression Of Snail & MMP-2 Of SW626 Cell Of Human Ovarian Cancerous Cell Line

Background and objectives:Ovarian cancer incidence in gynecological cancer ranks third in the mortality rate ranks first, with 60% to 90% of epithelial ovarian cancer (ovarian epithelial carcinoma, referred to as ovarian cancer).The rapid development of Epithelial ovarian cancer is likely to result in the transfer of lymphatic channels and blood line and the prognosis is poor, For the moment, ovarian cancer ranks first among all the gynecological malignancies in terms of mortality. Because it is hard to detect the ovarian cancer, when diagnosed, most are already at an advanced stage. with the treatment of surgery, radiotherapy and chemotherapy, some patients have completely recovered clinically, but about 70% of them will ultimately relapse. studies show that invasion and metastasis are not only important biological characteristics of malignant tumors, but also the leading cause for the death of those patients. In the course of invasion and spread of the tumor cells, adhesion molecules which involve in the interplay of cells form a basis for malignant tumors and the final target sites as well where many carcinogenic factors interact.Related research revealed that COX-2 contributes to the formation of tumors in the areas of cell apoptosis in gynecologic tumor, angiogenesis and tumor-infiltrating. Blocking COX-2 with NSAID has played an important role in the comprehensive treatment of gynecological tumors and the election of COX-2 inhibitors has become the latest research topic。In this study, the election of COX-2 inhibitor, (NS-398) will be applied to the highly metastatic cell line SW626 which is cultivated in vitro ovarian adenocarcinoma, observing its effects on the invasion and metastasis of ovarian cancer cell line. Exploring how the election of COX-2 inhibitor impacts the Snail protein of SW626 as well as MMP-2 expression with a purpose to probe into the molecular biological mechanism which may lead to the invasion and metastasis of tumor cells. Methods:Apply different concentrations of NS-398 (0,50, 100and 200 umol/L) to act on human ovarian cancerous cells, using MTT to determine how the concentration of NS-398 restrains the growth of SW626 cells in 24,48,72h time points, using real-time fluorescence quantitative PCR method to detect the expression NS-398 acts on the transcription level of Snail&MMP-2 mRNA expression in different concentrations (0,50,100& 200 umol/L) and different periods of time(24h,48h,72h). With different concentrations of COX-2 inhibitor (NS-398) 50,100&200 umol/L three concentrations and the time points 72h immunohistochemistry (SABC) Qualitative detection of Snail&MMP-2 at the protein level.Results:MTT method showed that:NS-398 inhibited the growth of SW626 was time and concentration dependent, and the control group (P<0.05). RT-PCR semi-quantitative results shows that NS-398 can greatly reduce Snail expression with a dose gradient descent and concentration-dependent (P<0.05),tendency which rises with increased concentration and prolonged time.NS-398 can reduce the expression of MMP-2 with duration protraction and concentration increase. (P<0.05). immunocyte chemistry result shows that,if the concentration of NS-398 increased,Snail and MMP-2 expreession will be decreased in protein level,the expression level based on the concentration,a significant difference will be found through microscope.Conclusion:Selective COX-2 inhibitors can inhibit the Snail & MMP-2 expression, and thereby can bring down Snail&MMP-2 mRNA and protein expression, which may be one of the mechanisms to reduce tumor cell invasion.