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合成Survivin小干扰RNA(small interfering RNA,siRNA)序列,用于转染人鼻咽癌细胞株CNE2,体外观察Survivin siRNA转染后CNE-2细胞Survivin mRNA、蛋白的表达、细胞增殖和凋亡情况。设计合成3段特异性Survivin siRNA,用siRNA转染鼻咽癌CNE-2细胞株,设置空白对照和阴性对照组,通过Real-time PCR检测CNE-2细胞的Survivin mRNA相对表达量,Western blotting检测Survivin蛋白的表达,流式细胞术及原位细胞凋亡检测转染后细胞凋亡情况。siRNA转染CNE-2细胞后,siRNA 1组和3组mRNA表达抑制率分别为(68.46±7.94)%、(49.44±3.78)%,siRNA 2组结果无显著差异(p>0.05)。蛋白相对表达量只有siRNA 1组降低,表达抑制率为(30.61±2.47)%,流式细胞术和原位细胞凋亡检测转染后CNE-2细胞数量明显降低,细胞凋亡比例增高。siRNA干扰沉默Survivin基因能有效抑制CNE-2细胞的增殖和诱导细胞凋亡。本研究为Survivin基因可能作为治疗鼻咽癌的一个靶点,为Survivin基因沉默可能是治疗鼻咽癌高效的途径提供体外实验支持。
Survivin siRNA was synthesized and transfected into human nasopharyngeal carcinoma cell line CNE2. The expression of Survivin mRNA and protein, proliferation and apoptosis of CNE-2 cells after Survivin siRNA transfection were observed in vitro . Three specific Survivin siRNAs were designed and synthesized. The CNE-2 cells were transfected with siRNA. Blank control group and negative control group were set up. The relative expression of Survivin mRNA in CNE-2 cells was detected by Real-time PCR. Survivin protein expression, flow cytometry and apoptosis in situ detection of apoptosis after transfection. After siRNA transfected into CNE-2 cells, the inhibitory rates of mRNA expression in group 1 and group 3 were (68.46 ± 7.94)% and (49.44 ± 3.78)%, respectively. There was no significant difference between siRNA group 2 and group 2 (p> 0.05). The relative expression of protein was only down-regulated in siRNA group 1 (30.61 ± 2.47)%. The number of CNE-2 cells and the percentage of apoptotic cells in transfected cells were significantly decreased by flow cytometry and in situ cell apoptosis assay. siRNA silencing Survivin gene can effectively inhibit CNE-2 cell proliferation and induce apoptosis. In this study, Survivin gene may serve as a target for the treatment of nasopharyngeal carcinoma, in vitro experimental support for Survivin gene silencing may be an effective way to treat nasopharyngeal carcinoma.