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Background: ATM plays an important role in response to DNA damage, while the roles of ATM in radiation-induced autophagy are still unclear in cervical cancer cells. @@@ Methods: Human cervical cancer cells, Hela, were used, and cell models with ATM(-/-) and MAPK14(-/-) were established by gene engineering. Western blot was implemented to detect protein expression. MDC staining and GFP-LC3 relocalization were used to detect autophagy. CCK-8 was used to detect cell viability. Radiosensitivity was analyzed by colony formation assays. Co-immunoprecipitation was used to detect the interaction between different proteins, and apoptosis was detected by flow cytometry. @@@ Results: After radiation autophagy was induced, illustrated by the increase of MAPLC3-II/MAPLC3-I ratio and decrease of p62, and phosphorylation of ATM simultaneously increased. ATM(-/-) cells displayed hypersensitivity but had no influence on IR-induced apoptosis. Then inhibitor of ATM, KU55933, ATM and MAPK14 silencing were used, and autophagy was induced by IR more than 200% in control, and only by 35.72%, 53.18% and 24.76% in KU55933-treated cells, ATM(-/-) and MAPK14(-/-) cells, respectively. KU55933 inhibited IR-induced autophagy by activating mTOR pathways. ATM silencing decreased the expression of MAPK14 and mTOR signals significantly. Beclin's bond to P13KIII and their interaction increased after IR, while in ATM(-/-) and MAPK14(-/-) cells this interaction decreased after R Both ATM and MAPK14 interacted with Beclin, while ATM(-/-) and MAPK14(-/-) cells showed no interaction. @@@ Conclusions: ATM could promote IR-induced autophagy via the MAPK14 pathway, the mTOR pathway, and Beclin/PI3KII complexes, which contributed to the effect of ATM on radiosensitivity.
- 出版日期2013-12
- 单位吉林大学
