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目的:应用hTERT特异性siRNA干扰舌癌Tca8113细胞hTERT基因的表达,探讨hTERT-siRNA联合顺铂对舌癌Tca8113细胞增殖和凋亡的影响。方法:体外化学合成针对hTERT的siRNA,通过阳离子脂质体将siRNA-hTERT1转染舌癌Tca8113细胞,RT-PCR检测hTERTmRNA表达水平,Western印迹检测其蛋白表达水平,四甲基偶氮唑蓝法(MTT)及流式细胞分析法分别测定hTERT-siRNA联合顺铂处理后对舌癌Tca8113细胞增殖及凋亡的影响。数据采用SPSS13.0进行方差分析。结果:靶向hTERT的siRNA可以有效抑制舌癌Tca8113细胞株hTERT的表达,hTERT-siRNA和顺铂均可抑制Tca8113细胞增殖并诱导其一定程度的凋亡。当联合应用靶向hTERT的siRNA和顺铂时,上述作用显著加强(P<0.05)。结论:体外化学合成的hTERTsiRNA可以有效抑制舌癌Tca8113细胞hTERT的表达,hTERT表达受抑制后,可增强顺铂作用舌癌Tca8113细胞的敏感性。
Objective: To investigate the effect of hTERT-siRNA combined with cisplatin on the proliferation and apoptosis of tongue cancer Tca8113 cells by using hTERT-specific siRNA to interfere with the expression of hTERT gene in tongue cancer Tca8113 cells. Methods: siRNA targeting hTERT was synthesized in vitro. SiRNA-hTERT1 was transfected into tongue cancer Tca8113 cells by cationic liposomes. The expression of hTERT mRNA was detected by RT-PCR. The protein expression was detected by Western blotting. (MTT) and flow cytometry were used to determine the effect of hTERT-siRNA combined with cisplatin on the proliferation and apoptosis of tongue cancer Tca8113 cells. Data were analyzed by ANOVA using SPSS13.0. Results: siRNA targeting hTERT effectively inhibited the expression of hTERT in tongue cancer Tca8113 cells. Both hTERT-siRNA and cisplatin inhibited the proliferation of Tca8113 cells and induced some degree of apoptosis. This effect was significantly enhanced when siRNAs targeting hTERT were combined with cisplatin (P <0.05). Conclusion: hTERTsiRNA synthesized in vitro can effectively inhibit the expression of hTERT in tongue cancer Tca8113 cells. After hTERT expression is inhibited, the sensitivity of cisplatin to Tca8113 cells can be enhanced.