顺铂处理卵巢癌细胞系OVCAR3引起Survivin表达差异的研究

Expression of survivin mRNA and protein in ovarian cancer cell line( OVCAR3) during cisplatin treatment

的 探讨顺铂(DDP)处理卵巢癌细胞系时Sur-vivin表达是否有变化,进而探讨Survivin表达与肿瘤细胞耐药之间的关系及其在肿瘤细胞获得性耐药及细胞凋亡耐受所起的重要作用。方法 以卵巢癌细胞系OVCAR3为材料,用MTT比色法检测细胞活性,流式细胞术检测细胞凋亡。RT-PCR检测Survivin mRNA表达差异,Western blot检测Survivin蛋白表达变化。结果 顺铂抑制卵巢癌细胞生长,具有时间和浓度依赖效应,即随时间延长和浓度增加,细胞生长抑制越强,细胞凋亡率也有同样的趋势,最高达31.6％。1mg·L^(-1)DDP处理卵巢癌细胞,Survivin mRNA及蛋白表达随作用时间的增长,分别在8 h和12 h达最高,随后又降低。结论 抗凋亡蛋白Survivin在卵巢癌细胞系中呈高表达,顺铂处理卵巢癌细胞,Survivin mRNA表达随作用时间延长而增加,这可能是卵巢癌细胞对化疗药物产生耐药性的重要因素之一。

AIM In the prestent study, the changes in expression levels of survivin messenger RNA(mRNA) and survivin protein in ovarin cancer cell line OVCAR3 during cisplatin (DDP) treatment were analyzed and compared with the occur-ance of programmed cell death with various concentration of DDP so to investigate the correlation between survivin expression and chemoresistance of cancer cells. METHODS MTT assay was used to analyze inhibition of cell growth. Flowcytome-try was used to analyze the changes of apoptosis index(AI) of OVCAR3 cell after having been treated with DDP at serials of concentration and time. RT-PCR and Western blot were used to analyze the changes in expression levels of survivin mRNA and protein. RESULTS Time- and dose- dependent effects on the growth inhibition with DDP treatment were observed. As time and doseage in- creased, the growth inhibition also increased. The apoptotic index showed similar trend. Survivin mRNA and protein increased within 8 h and 12 h, and reached peak level at 8 h and 12 h respectively then reduced after that time. CONCLUSION Expression of survivin mRNA and protein increases in OVCAR3 cells with DDP treatment suggesting that survivin may correlate with the chemoresis-tence of ovary malignant cells.