siRNA沉默Survivin基因对鼻咽癌CNE-2细胞凋亡的影响

Effect of siRNA Silencing Survivin Gene on Apoptosis of CNE-2 Cells in Nasopharyngeal Carcinoma

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。

Small interfering RNA （siRNA） sequence of Survivin gene was synthesized to transfect human nasopharyngeal carcinoma cell line CNE2. The Survivin mRNA, protein expression, cell proliferation and apoptosis of CNE-2 cells transfected with Survivin siRNA were observed. 3 Survivin siRNA segments with specificity were designed and synthesized, then siRNA transfected CNE-2 cell line. The blank control and negative control group were set, the relative expression of Survivin mRNA in CNE-2 cells was detected by PCR Real-Time, the protein expression of Survivin was detected by Western blotting, and the apoptosis of transfected cells was detected by flow cytometry and in situ cell apoptosis. After transfection of CNE-2 cells with siRNA, the inhibitory rates of mRNA expression in 1 group and 3 group were （68.46 ±7.94）%, （49.44±3.78）%, respectively, but the result of siRNA 2 group was not statistically significant （p 〉0.05）. The relative expression of protein was decreased only in siRNA 1 group, and the expression inhibition rate was （30.61 ±2.47）%. Flow cytometry and in situ cell apoptosis showed that CNE-2 cell number was significantly decreased and the percentage of apoptosis was increased after transfection. The interference of siRNA on silencing Survivin gene could effectively inhibit the proliferation of CNE-2 cells and induce their apoptosis. This study suggested that Survivin gene might be a target spot in the treatment of nasopharyngeal carcinoma, which provided in vitro experimental support for Survivin gene silencing might be an effective way for the treatment ofnasopharyngeal carcinoma.