目的 探讨肾癌786-O细胞Survivin基因对肾癌增殖能力的影响.方法 采用RNA干扰技术转染肾癌786-O细胞,靶向沉默Survivin基因表达,通过RT-PCR和Western blot实验检测沉默效率,并测定增殖细胞核抗原PCNA蛋白表达变化,通过MTS实验测定细胞增殖活性.结果 干扰组Survivin mRNA、Survivin蛋白及PCNA蛋白表达水平明显低于对照组和空白组,差异均有统计学意义(P＜0.05).对照组与空白组SurvivinmRNA、Survivin蛋白及PCNA蛋白相对表达量差异均无统计学意义(P＞0.05).3组786-O细胞增殖活力均呈 升高趋势,干扰组786-O细胞增殖活力明显低于对照组和空白组,其组间、时点间、组间·时点间交互作用差异均有统计学意义(P＜0.05).结论 RNA干扰Survivin基因能显著下调肾癌786-O细胞的Survivin基因表达,并降低细胞增殖活力,揭示出Survivin基因影响肾癌细胞恶性增殖的潜在分子机制.%Objective To investigate the effect of Survivin gene expression on human renal carcinoma cell 786-O proliferation.Methods RNA inteference was used to transfect human renal cancer cell line 786-O and silence Survivin gene expression.Expression of Survivin mRNA and protein were detected by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot assay,respectively.Change of PCNA protein expression was measured meanwhile.The proliferation of 786-O cells was evaluated by MTS assay.Results Compared to control siRNA/786-O and non-siRNA/786-O,Survivin siRNA/786-O displayed reduced Survivin expression of mRNA and protein (P＜ 0.05).Expression level of PCNA protein was also down-regulated significantly in Survivin siRNA/786-O cells (P＜0.05).No significant differences of Survivin mRNA,Survivin protein and PCNA protein were found between control siRNA/786-O and nonsiRNA/786-O(P ＞ 0.05).Proliferation activity of all three groups of 786-O cells showed increasing trend obviously.The level of siRNA/786-O cells proliferation was significantly lower than other groups.There were statistically significant differences between groups,at different time point,also analysis of interaction between groups with time point revealed statistical significance(P＜ 0.05).Conclusion RNA interference of Survivin gene can down-regulate expression of Survivin mRNA,Survivin protein and PCNA protein expression with decreasing the proliferation activity of cells.The present study revealed underlying influence of Survivin gene expression on malignant proliferation of renal cancer.
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