军事医学2016，Vol.40Issue(7)：537-540,4.DOI:10.7644/j.issn.1674-9960.2016.07.001
DNA依赖性蛋白激酶催化亚基在电离辐射诱发HeLa细胞自噬中的作用
The role of DNA-PKcs in autophagy induced by ionizing radiation
摘要
Abstract
Objective To investigate the effect of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) on autophagy induction by ionizing radiation ( IR ) .Methods The cell model of knocking-down DNA-PKcs expression was constructed by transfecting HeLa cells with a pSicoR-based lentivirus vector expressing DNA-PKcs specific shRNA .Cellular growth activity and radiosensitivity were detected by cell countingkit ( CCK)-8 assay.The expression of autophagy related proteins was detected by Western blotting hybridization .Autophagy was also detected by monitoring the autophagic marker green fluorescere protein ( GFP )-light chain 3 ( LC3 ) puncta per cell under an immunofluorescent microscope .Results A cellular model of knocking-down DNA-PKcs expression was successfully generated by transfecting the specific shRNA against DNA-PKcs.Depression of DNA-PKcs significantly decreased the growth activity of HeLa cells and increased the cellular sensitivity to ionizing radiation .Both the expression changes of P 62 and LC3 proteins and immunofluorescent GFP-LC3 puncta observation indicated that knocking-down DNA-PKcs prompted the induction of autophagy by ionizing radiation . Moreover, inactivation of DNA-PKcs led to a decreased phosphorylation of mammalian target of sirolimus ( Rapamycin, RAPA) ( mTOR) at S2481 site.Conclusion Depression of DNA-PKcs expression prompts the induction of autophagy by IR and cellular radiosensitivity .mTOR signaling may be involved in the regulation of autophagy processing by DNA-PKcs.关键词
DNA依赖性蛋白激酶催化亚基/电离辐射/HeLa细胞/细胞增殖/自噬/西罗莫司/哺乳动物西罗莫司靶蛋白( mTOR)Key words
DNA-PKcs/ionizing radiation/HeLa cell/cell proliferation/autophagy/sirolimus/mTOR分类
医药卫生引用本文复制引用
韩艳勤,王豫,刘晓丹,周平坤..DNA依赖性蛋白激酶催化亚基在电离辐射诱发HeLa细胞自噬中的作用[J].军事医学,2016,40(7):537-540,4.基金项目
国家自然科学基金资助项目 （）
