沉默Chk1基因对姜黄素诱导胃癌细胞SGC7901凋亡敏感度的影响
Effect of Chk1 Gene Silencing on Curcumin-induced Apoptosis Susceptibility of Human Gastric Cancer SGC7901 Cells
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摘要: 目的 探讨siRNA沉默Chk1基因表达对姜黄素诱导胃癌细胞SGC7901凋亡和细胞周期的影响，评价其作为姜黄素治疗胃癌增敏靶点的有效性。 方法 采用RNAi技术在胃癌细胞SG-7901中将Chk1基因沉默，采用Western blot检测转染前后Chk1蛋白表达的变化，采用流式细胞术检测Chk1基因沉默对姜黄素诱导胃癌细胞凋亡及细胞周期变化的影响。 结果 转染Chk1 siRNA后，胃癌细胞SGC7901中Chk1蛋白表达受抑制，明显低于对照组(P＜0.05)。FCM检测结果显示，si-Chk1组较空白对照组G2/M期百分比有所降低(P＜0.05)，si-Chk1+Curcumin组G2/M期比例明显低于Empty vector+Curcumin组(P＜0.05)。siRNA沉默Chk1基因使姜黄素诱导的细胞凋亡率由(14.7±1.1)%上升到(28.9±1.8)%。 结论 siRNA沉默Chk1基因可明显消除G2/M期阻滞，并显著增强姜黄素诱导胃癌细胞SGC-7901凋亡的敏感度，提示Chk1可作为姜黄素治疗胃癌的有效增敏靶点。Abstract: Objective To investigate the effect of siRNA targeting checkpoint kinase 1 (Chik1) on curcumin-induced apopotosis and cell cycle of human gastric cancer SGC7901 cells,so as to evaluate the role of Chk1 as a therapeutic target to sensitize human gastric cancer to curcumin.Methods The SGC7901 cells were transfected with siRNA-Chk1.The protein expression of Chk1 was detected by Western blotting.Cell cycle phase distribution and cell apoptosis were determined by flow cytometry (FCM). Results Compared with the control group,the Chk1 protein expression was significantly reduced in siRNA-Chk1 transfection group (P＜0.05).Further investigation revealed that the percentage of the G2/M phase cells of si-Chk1 group was lower of than that of the control group (P＜0.05) and the percentage of the G2/M phase cells was lower of than that of Empty vector+Curcumin group (P＜0.05).Transfection of si-Chk1 led to increased apoptotic rate from (14.7±1.1)% to (28.9±1.8)%. Conclusion Transfection of si-Chk1 decreased G2/M arrest and sensitized SGC7901 cells to curcumin-induced apoptosis,suggesting that Chk1 could be a potential therapeutic target to sensitize human gastric cancer to curcumin.
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