The Invitro Study Of RNA Interference Targeted Survivin Gene Treated The Epithelial Ovarian Carcinoma

Ovarian cancer is one of three malignancies of gynecological , Itassociated with the highest incidence and mortality rate among allgynecological neoplasms . Human common epithelial ovarian carcinomascomprise 59% of all ovarian tumors and up to 90% of primary ovarianmalignancies. At present , the main therapy of human epithelial ovarianis surgery, combined with chemotherapy .But , Because its mostly werediscovered in late , the surgery effect was poor or had no opportunity toreceive operating at all , and the serious resistant to chemotherapy drugs ,epithelial ovarian carcinomas had the highest fatality rate. |It is badlyneed to seek a new therapeutic tool. RNA interference (RNAi) was a new approach of silence geneexpression which was discovered recently . It has been widely used invarious species , such as tumors gene therapy ,antivirus infection,filter ofgene-drugs and so on . As others gene therapy technic , the most difficult inusing is how to choice its target gene . Survivin, a novel structurally uniqueinhibitor of apoptosis (IAP), is undetectable in terminally differentiatedadult tissues, but becomes notably expressed in most common humancancers . The High expression of survivin gene in melanoma play animportant role in tumor cells apoptosis and generation of tumors bloodvesseles ,It also related to tumors resistance to chemotherapy andradiotherapy .It was acknowledged as the ideal target of gene therapy. In this study, we detected the expression of survivin in epithelialovarian cancers drug resistance cell line SKOV3/ADM and its parental cellline SKOV3, sought to determine whether RNAi-mediated silencing ofsurvivin expression could effectively induce apoptosis of these two celllines. At the same time , reversal of drug resistance in SKOV3/ADM afterRNA interference silencing survivin.Try to find new ideology and establishsteady laboratory basis of ovarian cancers gene therapy . We explored thatsilencing of survivin associated with RNAi-based strategy should be furtherexplored as a potentially efficacious way to treat ovarian cancer . PART Ⅰ EXPRESSION AND SIGNIFICANCE OF SURVIVIN GENE IN OVARIAN CANCER CELL LINES SKOV3 AND SKOV3/ADM Objective: The study was designed to examine the expression ofsurvivin gene in ovarian epithelial carcinoma cell line SKOV3 and its drugresistance cell line SKOV3/ADM , then discussed its significance . Methods : We cultured the ovarian epithelial cancer cell lines SKOV3and SKOV3/ADM . Semi-quantitative RT-PCR was used to detectedexpression of survivin mRNA in these two cell lines , expression ofsurvivin gene protein was measured by Western blot andimmunofluorescence . Results: 1.Semi-quantitative RT-PCR showed that mRNA transcription ofsurvivin gene was highly expressed in cell lines SKOV3 and SKOV3/ADMat comparatively ratio of 75.3% and 99.1%. 2.According to results of Western blot the comparatively expressionrate of survivin protein in these two cell lines were 76.3% and 99.8%. 3.After immunofluorescence staning 41.64±3.42 and 59.34±5.39fluorescent cells percent were detected in cell lines SKOV3 andSKOV3/ADM, satistatic value P<0.05. Conclusions: survivin gene was over-expressed in ovarian epithelialcarcinoma cell lines SKOV3 and SKOV3/ADM, The level of survivin geneexpressed in SKOV3/ADM obvisously difference compared with theexpression in its parental cell line SKOV3 , suggested that survivin genemay play an important role in the development of ovarian cancer and it wasone of the critical factors in ovarian cancer cells chemotherapy resistance. PART ⅡTHE STUDY OF RNAI-MEDIATED SILENCING EXPRESSION OF SURVIVIN GENE IN OVARIAN CANCER CELL LINES SKOV3 AND SKOV3/ADM AND ITS EFFECTS ON CELLS APOPTOSIS Objective: In this study we apply siRNA directed against survivingene to inhibit expression of survivin gene in ovarian cancer cell linesSKOV3 AND SKOV3/ADM, discussed effects that RNA interferenceagainst survivin gene decreasing expression of survivin gene and increasingthe apoptosis of cells. Methods: 1.PshRNA-survivin was transfected in SKOV3/ADM ce...