电离辐射对Beclin1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

doi:10.13481/j.1671-587x.20150608

Abstract

Abstract:

Objective To establish the MCF-7 cell models of Beclin 1 over-and low-expressions, and to detect the autophagic and apoptotic changes after 4 Gy irradiation, and to explore their molecular regulation mechanisms. Methods MCF-7,MCF-7+4Gy,MCF-7-Beclin 1+4Gy and MCF-7-Belcin 1 RNAi+4Gy groups were set up. Molecular biology method was used to construct Beclin 1 over-expression vector pcDNA3.1-Beclin 1, and to estabilish the Beclin 1 over-and low-expression cell models. After the cells were irradiated with 4 Gy, the autopahgic cell percentages were measured by fluorescence microscope with MDC staining, the apoptotic cell percentages were measured by FCM with AnnexinⅤ-FITC and PI staining, and the expressions of Beclin1, P53, Bcl-2 and Bax proteins were measured by Western blotting method. Results Compared with MCF-7 group, the autophagic and apoptotic cell percentages in MCF-7+4 Gy, MCF-7 Beclin 1+4 Gy and MCF-7-Beclin 1 RNAi+4 Gy groups were significantly increased (P<0.05 or P<0.001 ), especially in MCF-7 Beclin 1+4 Gy group which was significantly higher than those in MCF-7+4 Gy (P<0.05); while there was significant difference in the necrotic cell percentages between various groups. After 4 Gy irradiation, compared with MCF-7 group, the expression levels of Beclin 1, P53 and Bax proteins in MCF-7+4 Gy and MCF-7-Beclin 1+4 Gy groups were increased, but the expression levels of Bcl-2 protein were decreased, especially in MCF-7-Beclin 1+4 Gy group. Conclusion The MCF-7 cell models of Beclin 1 over-and low-expressions are successfully established, and ionizing radiation could induce the autophagy and apoptosis of MCF-7 cells, which is more obvious in Beclin 1 over-expression MCF-7 cells. Beclin 1 can activate P53, inhibit Bcl-2 and activate Bax,which forms the regulation of autophagy and apoptosis by P53.

Key words: autophagy, aopotosis, ionizing radiation, Beclin 1, P53

CLC Number:

R818

ZHANG Jing, ZHAO Dali, XIE Zhongwei, LIU Yanjun, LI Zhijun, LI Yan, GONG Shouliang, QI Yali. Effects of ionizing radiation on autophagy and apoptosis in MCF-7 cells with Beclin 1 over-and low-expressions and their regulating mechanisms[J].Journal of Jilin University Medicine Edition, 2015, 41(06): 1139-1143.

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Effects of ionizing radiation on autophagy and apoptosis in MCF-7 cells with Beclin 1 over-and low-expressions and their regulating mechanisms

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