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目的:探讨c-raf-1基因反义寡核苷酸转染对人卵巢癌SKOV3细胞增殖的抑制作用及其分子机制。方法:实验分3组:正常对照组、c-raf-1正义治疗组、c-raf-1反义治疗组。脂质体转染后不同时间分别进行M TT试验、集落形成试验、荧光显微镜检测、蛋白质荧光强度测定,以观察不同条件处理后各组细胞的增殖、凋亡、蛋白质表达有无差别。结果:反义治疗组与正义治疗组比较:转染72h OD值分别0.272与1.307(P<0.05);克隆形成率分别为30.6%与75.0%(P<0.05);凋亡率分别为19.7%与9.2%(P<0.05),同时明显地下调P74蛋白质的表达水平(P<0.05)。结论:c-raf-1反义寡核苷酸对人卵巢癌SKOV3细胞的增殖有明显的抑制作用。
Objective: To investigate the inhibitory effect of c-raf-1 antisense oligonucleotide transfection on the proliferation of human ovarian cancer cell line SKOV3 and its molecular mechanism. Methods: The experiment was divided into three groups: normal control group, c-raf-1 sense therapy group and c-raf-1 antisense therapy group. M TT test, colony formation assay, fluorescence microscopy and protein fluorescence intensity were performed at different times after lipofectamine transfection to observe the proliferation, apoptosis and protein expression of the cells in different conditions. Results: The OD value of antisense treatment group was 0.272 and 1.307 (P <0.05) at 72h after transfection, respectively. The colony formation rates were 30.6% and 75.0% respectively (P <0.05), and the apoptotic rates were respectively 19.7% And 9.2% (P <0.05), respectively. At the same time, P74 protein expression was significantly down-regulated (P <0.05). Conclusion: The antisense oligonucleotide c-raf-1 can significantly inhibit the proliferation of human ovarian cancer cell line SKOV3.