Survivin siRNA沉默对非小细胞肺癌细胞生物学的影响

The biological effect of Survivin on NSCLC cell line by siRNA targeting Survivin

目的探讨凋亡抑制蛋白(Survivin)对非小细胞肺癌(NSCLC)H1299细胞株的生物学影响。方法通过脂质体介导将Survivin基因siRNA瞬时转染H1299细胞。采用RT-PCR及Western blot检测转染后H1299细胞内Survivin基因的表达水平。MTT比色法、流式细胞术(FCM)及Western blot观察细胞增殖、凋亡、周期及Caspase-9蛋白表达变化情况。结果 RT-PCR及Western blot证实,siRNA沉默的H1299细胞中Survivin mRNA、蛋白的表达水平较未转染组明显降低(t=10.970,P=0.000;t=9.192,P=0.003)。MTT、细胞凋亡及周期结果表明,siRNA沉默的H1299细胞其增殖能力明显减弱、细胞凋亡明显升高,G2/M期阻滞,Caspase-9蛋白表达增加(t=8.162,P=0.001;t=10.800,P=0.000;t=9.192,P=0.003)。结论 Survivin参与NSCLC细胞增殖、凋亡、周期这一生物学过程。

Objective To investigate the effect of inhibitor of apoptosis proteins （Survivin） on biological effect of human lung cancer cell line. Methods Survivin siRNA was transiently transfected into non small cell lung carcinoma （NSCLC） cell H1299 by liposome-mediated method and was detected by RT-PCR and western blot. MTT assay, cell apoptosis, cell cycle and western blot were also conducted as to the influence of the down-regulated expression of Survivin that might be found on H1299 cells biological effect. Results RT-PCR and Western blot showed that H1299 cell transfected Survivin siRNA had a lower relative expressive content than normal cell （t=10.970, P=0.000; t=9.192, P=0.003）. MTT assay, cell apoptosis, cell cycles demonstrated that H1299 cell transfected Survivin siRNA had a lower survival fraction, higher cell apoptosis, more percentage of the G2/M phases （t=8.162, P=0.001;t=10.800, P=0.000）. Western blot showed that the relative content of Caspase-9 protein was increased in H1299 cell transfected Survivin siRNA compared with H1299 cell untransfected Survivin （t=9.192, P=0.003）. Conclusion Survivin may involved in the biological processes of lung cancer cell proliferation, apoptosis, and cell cycle.