SUMO Expression and Regulation in Oxidative Stress in Cultured Human Lens Epithelial Cells
- VernacularTitle:SUMO蛋白在体外培养的人晶状体上皮细胞内的表达及对氧化应激的调控作用
- Author:
Xiao HAN
;
Xinling WANG
;
Di WU
;
Jinsong ZHANG
;
Qichang YAN
- Publication Type:Journal Article
- Keywords:
small ubiquitin-related modifiers;
human lens epithelial cells;
cataract;
high glucose;
oxidative stress
- From:
Journal of China Medical University
2015;(3):193-198
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the expression of small ubiquitin?related modifiers(SUMO)protein in normal cultured human lens epithelial cells(SRA01/04)and discuss regulation effects of SUMO protein on oxidative stress induced by high glucose. Methods The expression and local?ization of SUMO 1,2/3,4 was detected in normal cultured SRA01/04 cells through immunocytochemistry. The mRNA expression levels of SUMO 1?4 were examined by RT?PCR after the SRA01/04 cells treated with high glucose media at different concentrations and time points. Samples were grouped by medium concentrations(glucoses 5.5 mmol/L,12.5 mmol/L,25 mmol/L,50 mmol/L respectively for 24 h)and by treatment time(0 h, 6 h,12 h and 24 h respectively). After highly efficient transfection of GFP?SUMO2 into SRA01/04 cells,the survival and apoptotic rates of transfect?ed and un?transfected cells treated with high glucose was detected by CCK8 method and AV/PI double staining flow cytometry. Results The immu?nocytochemistry results showed that SUMO1,2/3,4 proteins were mainly located in the nucleus of SRA01/04 cells and part of SUMO2/3 was in the cytoplasm. RT?PCR results showed that compared with the low?glucose group,the mRNA expression of SUMO1?4 was increased along the increas?ing glucose concentration in the high?glucose group(P<0.05). Compared with 0 h,the mRNA expression of SUMO1?4 was enhanced at 6 h,12 h and 24 h(P<0.05)in the high?glucose group treated at 50 mmol/L concentration. Compared with the un?transfected cells,the survival rate was in?creased and the apoptotic rate was decreased in GFP?SUMO2 transfected cells in oxidative stress induced by high glucose(P<0.05). Conclusion SUMO protein was positively expressed in SRA01/04 cells and the expression of SUMO mRNA was affected by oxidative stress induced by high glu?cose.
