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目的探讨体外转染靶向生存素的小干扰RNA(siRNA)是否能够增强厄洛替尼对肺腺癌细胞的增殖抑制、凋亡诱导作用,以及厄洛替尼是否能够抑制肺腺癌细胞生存素基因的表达,并探讨其体外作用机制。方法体外培养人肺腺癌A549细胞,转染靶向生存素的siRNA和厄洛替尼两种处理因素单独或联合应用于A549细胞,流式细胞技术检测各组A549细胞早期凋亡情况;噻唑蓝比色法检测A549细胞增殖抑制情况;免疫细胞化学染色法观察各组A549细胞生存素蛋白表达的变化;免疫蛋白印迹法检测各组A549细胞生存素蛋白表达水平。结果生存素siRNA转染和厄洛替尼两者单独或联合应用于人肺腺癌.A549细胞作用48 h后,A549细胞的早期凋亡率均明显增加,都对细胞增殖有明显的抑制作用,并能降低细胞内生存素蛋白的表达水平,与空白对照组比较差别有统计学意义(P<0.05),两者联合应用较单独应用对A549细胞增殖的抑制作用更显著、早期凋亡更明显、对生存素蛋白表达的抑制作用更强(均P<0.05)。结论应用siRNA沉默生存素表达是提高体外厄洛替尼对人肺腺癌A549细胞增殖抑制、凋亡诱导及生存素蛋白表达抑制作用的有效途径。
OBJECTIVE: To investigate whether small interfering RNA (siRNA) targeted to survivin transfected in vitro can enhance the inhibitory effect of erlotinib on the proliferation of lung adenocarcinoma cells and the induction of apoptosis, and whether erlotinib can inhibit the survival of lung adenocarcinoma cells Prime gene expression, and explore its mechanism in vitro. Methods Human lung adenocarcinoma A549 cells were cultured in vitro and transfected with survivin siRNA and erlotinib. A549 cells were treated with A549 cells alone or in combination. Flow cytometry was used to detect the early apoptosis of A549 cells. The inhibition of proliferation of A549 cells was detected by blue colorimetric assay. The expression of survivin protein in A549 cells was observed by immunocytochemical staining. The expression of survivin in A549 cells was detected by Western blotting. Results Survivin siRNA transfection and erlotinib alone or in combination with human lung adenocarcinoma A549 cells 48 h after the apoptosis rate of A549 cells were significantly increased both have a significant inhibitory effect on cell proliferation , And decreased the expression of survivin protein in cells compared with the blank control group (P <0.05). The combination of the two drugs had a more significant inhibitory effect on the proliferation of A549 cells compared with the control alone, and the early apoptosis was more Obviously, the inhibition of Survivin protein expression was stronger (all P <0.05). Conclusion The application of siRNA to silence survivin expression is an effective way to increase the inhibitory effect of erlotinib on human lung adenocarcinoma A549 cell proliferation, apoptosis induction and survivin protein expression in vitro.