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目的:研究特异性siRNA对胃癌细胞的抑制作用,以及探讨采用RNA干扰技术下调Bcl-2基因能否增强SGC-7901胃癌细胞对紫杉醇的敏感性。方法:通过脂质体HiperFect将Bcl-2 siRNA转染入胃癌细胞SGC-7901;采用RT-PCR和蛋白印迹法检测Bcl-2 siRNA转染前后胃癌细胞Bcl-2基因表达的变化;用Annexin Ⅴ法及细胞周期分析法检测细胞凋亡;用MTS法观察细胞对药物紫杉醇敏感性的影响。结果:Bcl-2 siRNA组较空白对照组明显抑制Bcl-2 mRNA的表达水平,抑制率为(87.6±5.3)%,P=0.001;Bcl-2 siRNA组较空白对照组明显下调Bcl-2蛋白表达水平,抑制率为(85.7±3.6)%,P=0.003。Bcl-2 siRNA组、紫杉醇组和紫杉醇+Bcl-2 siRNA细胞总凋亡率分别为49.00%、46.10%和66.00%,联合用药组细胞凋亡率最强。Bcl-2siRNA+紫杉醇联合用药对SGC-7901细胞周期分析结果显示,联合用药组凋亡率为48.00%,Bcl-2 siRNA组和紫杉醇组分别为9.03%和21.50%。Bcl-2 siRNA组、阴性对照组和空白对照组的IC50分别(7.25±0.22)、(54.45±0.82)和(68.9±0.91)μg/L,Bcl-2 siRNA转染胃癌细胞SGC-7901对药物紫杉醇更敏感,其IC50值比阴性siRNA转染组降低86.7%,即对紫杉醇敏感性增加7.25倍。结论:靶向Bcl-2 siRNA可明显下调靶基因Bcl-2表达,促进SGC-7901细胞凋亡并能提高细胞对紫杉醇敏感性,为胃癌治疗提供新的策略。
OBJECTIVE: To investigate the inhibitory effect of specific siRNA on gastric cancer cells and to investigate whether the down-regulation of Bcl-2 gene by RNA interference can enhance the sensitivity of SGC-7901 gastric cancer cells to paclitaxel. Methods: Bcl-2 siRNA was transfected into SGC-7901 gastric cancer cells by lipofection HiperFect. The expression of Bcl-2 gene in gastric cancer cells before and after Bcl-2 siRNA transfection was detected by RT-PCR and Western blotting. Apoptosis was detected by the method of cell cycle analysis and the effect of the cells on paclitaxel sensitivity by MTS method. Results: Compared with the blank control group, the Bcl-2 siRNA group significantly inhibited the expression of Bcl-2 mRNA (87.6 ± 5.3)%, P = 0.001; Bcl-2 siRNA group significantly decreased Bcl-2 protein The expression rate was (85.7 ± 3.6)%, P = 0.003. The total apoptotic rates of Bcl-2 siRNA group, paclitaxel group and paclitaxel + Bcl-2 siRNA group were 49.00%, 46.10% and 66.00%, respectively. The apoptosis rate of combination group was the strongest. Bcl-2siRNA + paclitaxel combined SGC-7901 cell cycle analysis showed that the apoptosis rate was 48.00%, Bcl-2 siRNA group and paclitaxel group were 9.03% and 21.50% respectively. The IC50 of Bcl-2 siRNA group, negative control group and blank control group were (7.25 ± 0.22), (54.45 ± 0.82) and (68.9 ± 0.91) μg / L respectively. The Bcl-2 siRNA transfected SGC-7901 Paclitaxel was more sensitive and its IC50 value was 86.7% lower than that of negative siRNA transfection group, which was 7.25 times more sensitive to paclitaxel. CONCLUSION: Targeting Bcl-2 siRNA can significantly down-regulate the expression of Bcl-2, promote the apoptosis of SGC-7901 cells and increase the sensitivity of paclitaxel to SGC-7901 cells. It provides a new strategy for the treatment of gastric cancer.