线粒体靶向KillerRed增强辐射诱导HeLa细胞自噬作用及其机制

doi:10.13481/j.1671-587x.20200405

Abstract

Abstract: Objective: To explore the enhancement of mitochondira-targeted KillerRed(mtKR) on the mitochondrial dysfunciton and autophagy caused by radiation in the HeLa cells, and to clarify the relative molecular mechanisms. Methods: After mitochondria-targeted expression vectors Pink-mtKR were transfected into the HeLa cells, the cells were irradiated by visible light and 4 Gy X-ray. The cells were divided into control, empty vector, Pink1-mtKR, control + 4 Gy X-ray irradiation, empty vector + 4 Gy X-ray irradiation and Pink1-mtKR+4 Gy X-ray irradiation groups. After the cells were irradiated with X-ray for 24 h, the mitochondrial membrane potentials (MMP) and the autophgic rates were detected by flow cytometry, the levels of mitochondrial respiratory chain complex Ⅰ and Ⅲ were measured by biochemical assay,and the expression amounts of P62, Pink1, parkin and Tom20 proteins were measured by Western blotting method. Results: After Pink1-mtKR plasmids were transfected into the HeLa cells, the cells were irradiated by visible light and 4 Gy X-ray; compared with control group, the MMP and the levels of mitochondrial respiratory chain complex Ⅰand Ⅲ in Pink1-mtKR group were significantly decreased (P<0.05), and the autohpagic rate was significantly increased (P<0.05); the MMP and the levels of mitochondrial respiratory chain complex Ⅰ and Ⅲ in Pink1-mtKR + 4 Gy X-ray irradiation group were decreased obviously (P<0.05), and the autohpagic rate was significantly increased(P<0.05). Compared with control group, the expression amounts of Pink1 and parkin proteins in total protein of the cells in Pink1-mtKR group and Pink1-mtKR+4 Gy X-ray irradiation group had no obvious changes, but the expression amounts of the P62 protein was increased; the expression amounts of Tom20, Pink1 and parkin in mitochondrial protein in Pink1-mtKR group and Pink1-mtKR+4 Gy X-ray irradiation group were increased obviously. Conclusion: MtKR may enhance the mitochondrial dysfunction and autohpagy caused by radiation, and its mechamism may be related to the regulation of Pink1/parkin autophgy pathway.

Key words: mitochondrion, reactive oxygen species, radiation, autophagy, mitochondrion-targeted KillerRed

CLC Number:

Q506

YU Lei, WANG Ce, HAN Bing, LI Xin, HAN Yuchen, SUN Yuying, GUO Xiangshu, LIU Weiwu, WANG Zhicheng. Enhancement of mitochondria-targeted KillerRed in autophagy caused by radiation in HeLa cells and its mechanism[J].Journal of Jilin University(Medicine Edition), 2020, 46(04): 693-698.

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Enhancement of mitochondria-targeted KillerRed in autophagy caused by radiation in HeLa cells and its mechanism

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