Effect ofα1-antitrypsin Z variant overexpression on autophagy regulation
10.3969/j.issn.1001-1978.2014.07.009
- VernacularTitle:α1抗胰蛋白酶突变体ATZ过表达增强细胞自噬水平
- Author:
Na ZHU
;
Lijie FENG
;
Haiping WANG
;
Yujun SHEN
;
Yuxian SHEN
- Publication Type:Journal Article
- Keywords:
α1-antitrypsin Z variant;
autophagy;
p62;
LC3;
Beclin1;
cytotoxicity
- From:
Chinese Pharmacological Bulletin
2014;(7):921-925
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate the effect of α1-anti-trypsin Z variant (ATZ)overexpression on cell autoph-agy.Methods HEK 293T cells were transfected with pcDNA3.1 zeo+/ATM or pcDNA3.1 zeo+/ATZ,e-qual amount of empty vector was used as control.Cells were treated with NH4Cl for 4 hours and processed for detecting ATZ,LC3 and p62 by immunoblot.Mean-while ,expression and intracellular localization of ATZ, LC3 in 293 T cells were observed with double labeled immunofluorescence.The mRNA levels of autophagy-related genes were measured by real-time PCR.Immu-nohistochemistry was used to observe the morphology of ATZ-positive cells.Results Compared with the control,higher LC3Ⅱ levels and LC3 puncta were observed in ATZ transfected cells.Meanwhile,the levelsof p62 were decreased in ATZ transfected cells,andreversed by NH4 Cl (25 mmol·L -1 )treatment.Overexpression of ATZ increased the mRNA levels of Atg5and Atg12,but had no obvious influence on Beclin1.ATZoverexpressing cells presented abnormal morphologies.The nuclei became reduced,condensed,and even disappeared in ATZpositive cells.Conclusion ATZ overexpression increases autophagy activity whichmay be related to increasing Atg5 and Atg12 levels.
