电离辐射对Beclin 1过表达和低表达MCF-7细胞模型细胞自噬和凋亡的影响及其调控机制

发布时间：2018-11-12 07:43

【摘要】：目的:建立Beclin 1过表达和低表达MCF-7细胞模型,检测4Gy照射后细胞自噬和凋亡的变化,探讨Beclin 1的分子调控作用机制。方法:实验分为MCF-7组、MCF-7+4Gy组、MCF-7-Beclin1+4Gy组和MCF-7-Belin1RNAi+4 Gy组。利用分子生物学方法构建Beclin 1过表达载体pcDNA3.1-Beclin 1,并建立Beclin 1过表达和低表达细胞模型。细胞经4 Gy照射后,采用MDC染色荧光显微镜观察自噬细胞百分比,AnnexinⅤ-FITC和PI染色流式细胞术检测凋亡细胞百分比,Western blotting法检测Beclin 1、P53、Bcl-2和Bax蛋白表达。结果:与MCF-7组比较,MCF-7+4Gy组、MCF-7-Beclin 1+4Gy组和MCF-7-Beclin 1RNAi+4Gy组自噬和凋亡细胞百分比均明显升高(P0.05或P0.01),以MCF-7-Beclin 1+4Gy组升高最明显,且高于MCF-7+4Gy组(P0.05);各组坏死细胞百分比无明显差异。4Gy照射后,MCF-7组和MCF-7-Beclin 1组细胞中Beclin 1、P53和Bax蛋白表达水平均升高,Bcl-2蛋白表达水平降低,且以MCF-7-Beclin 1组最为明显。结论:成功建立Beclin 1过表达和低表达MCF-7细胞模型,电离辐射能够诱导其细胞自噬和凋亡,且对Beclin 1过表达细胞作用更明显。Beclin1通过激活P53,进而抑制Bcl-2和激活Bax,从而形成以P53为中心的自噬与凋亡分子调控机制。
[Abstract]:Aim: to establish Beclin 1 overexpression and low expression MCF-7 cell model, to detect the changes of autophagy and apoptosis after 4Gy irradiation, and to explore the molecular regulatory mechanism of Beclin 1. Methods: the experiment was divided into MCF-7 group, MCF-7 4Gy group, MCF-7-Beclin1 4Gy group and MCF-7-Belin1RNAi 4 Gy group. Beclin 1 overexpression vector pcDNA3.1-Beclin 1 was constructed by molecular biological method, and Beclin 1 overexpression and low expression cell models were established. After 4 Gy irradiation, the percentage of autophagy cells was observed by MDC staining fluorescence microscope, and the percentage of apoptotic cells was detected by Annexin 鈪，

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