消岩汤含药血清介导下Survivin siRNA对人肺腺癌A549细胞增殖及凋亡的影响

Effect of Survivin siRNA on human lung adenocarcinoma A549 cell proliferation and apoptosis mediated by drug-containing serum of Xiaoyan decoction

关键词:  消岩汤  Survivin siRNA  A549  细胞增殖  凋亡

Key words:  Xiaoyan decoction  Survivin siRNA  A549  proliferation  apoptosis

摘要:

[目的] 分析消岩汤含药血清介导下Survivin siRNA对人肺腺癌A549细胞增殖及凋亡的影响，探究其抑瘤机制。[方法] 在Invitrogen公司的网上设计系统中设计靶向Survivin基因的siRNA序列，并将其转染A549细胞，应用噻唑蓝（MTT）法检测消岩汤以及转染组对A549细胞生长抑制的情况，免疫组化法观察A549细胞Survivin蛋白的表达情况，流式细胞术检测各组细胞凋亡情况。[结果] MTT法显示消岩汤具有抑制A549细胞生长的作用，其作用具有一定的时间依赖性，转染中药组的抑制率显著高于转染对照组及非转染中药组。经免疫组化法检测，转染对照组对Survivin蛋白表达的抑制率明显高于非转染中药组，但显著低于转染中药组。流式细胞术检测结果显示消岩汤及重组质粒以诱导细胞晚期凋亡为主，转染中药组A549细胞凋亡率显著高于其他各组。[结论] 消岩汤可诱导A549细胞的凋亡，联合应用转染靶向Survivin基因的重组质粒可增强其体外诱导A549细胞凋亡的作用。

Abstract:

[Objective] To analyze effect of Survivin siRNA on human lung adenocarcinoma A549 cell proliferation and apoptosis mediated by drug-containing serum of Xiaoyan decoction and probe into its mechanism of tumor suppression.[Methods] The siRNA targeting survivin gene sequence was designed by Invitrogen Company's online design system, and A549 cells were transfected. The growth inhibition of A549 cell by Xiaoyan decoction group and transfection group were detected by MTT method, the expression of Survivin protein of A549 cells was observed immunohistochemical method. Apoptosis of cells was detected by flow cytometry.[Results] MTT method showed that the effect of A549 cell growth was inhibited by the Xiaoyan Decoction and its function has certain time dependence. The inhibition rate of the transfection TCM group was significantly higher than that of the transfection control group and non transfection TCM group. Detected by immunohistochemical method, the inhibition rate of Survivin protein expression in transfected control group was significantly higher than that in non transfection TCM group, but significantly lower than transfection TCM group. Flow cytometry test results show that Xiaoyan Decoction and recombinant plasmid is given priority to inducing cell apoptosis in advanced stage, A549 cell apoptosis rate of transfection TCM group was significantly higher than other groups.[Conclusion] Xiaoyan Decoction can induce apoptosis of A549 cells. Combined the use of transfection targeting Survivin gene recombinant plasmid can enhance the effect of the apoptosis induction of A549 cells in vitro.