Expression of long-chain non-coding RNA-P21 in hydrogen peroxide induced human lens epithelial cells damage
10.3760/cma.j.cn115989-20221109-00522
- VernacularTitle:长链非编码RNA-P21在过氧化氢诱导的人晶状体上皮细胞损伤中的表达
- Author:
Xiaoming DONG
1
;
Yuxuan LIU
;
Liyang JI
;
Jing WANG
;
Jinsong ZHANG
Author Information
1. 沈阳爱尔卓越眼科医院 爱尔集团眼科医院集团白内障与人工晶状体研究所 沈阳爱尔眼科精准医疗研究所 中国医科大学附属第四医院眼科 中国医科大学眼科医院 辽宁省晶状体研究重点实验室，沈阳 110000
- Keywords:
Lens epithelial cells;
Long-chain non-coding RNA-p21;
Proliferation;
Apoptosis;
Oxidative stress
- From:
Chinese Journal of Experimental Ophthalmology
2024;42(3):232-239
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To detect the changes in the biological activity and expression of long-chain non-coding RNA-p21 (lncRNA-p21) in human lens epithelial cells HLE-B3 damage induced by hydrogen peroxide.Methods:HLE-B3 cells were divided into normal control group and hydrogen peroxide group, which were cultured in normal culture medium and culture medium containing 200 μmol/L hydrogen peroxide for 24 hours, respectively.Cell viability was determined by MTS colorimetric method.Cellular reactive oxygen species (ROS) level was detected using ROS assay kits.Cell apoptosis was tested by flow cytometry.Cell Caspase-3 activity was detected using Caspase-3 assay kit.Expressions of Bax and Bcl-2 proteins related to cell apoptosis were determined by Western Blot.Cell cycle distribution was determined by flow cytometry.Cell proliferation ability was detected by EDU proliferation assay kit.The expression of lncRNA-p21 in cells was detected by real time fluorescence quantitative polymerase chain reaction (PCR).The localization of lncRNA-p21 in cells was detected by fluorescence in situ hybridization.Results:The ROS content of cells in hydrogen peroxide group was (4.65±0.38), significantly higher than (1.00±0.01) of normal control group, and the difference was statistically significant ( t=16.66, P<0.05).Compared with the normal control group, the cell apoptosis rate was significantly increased, the activity of Caspase-3 was enhanced, and the relative expression of Bax was significantly increased in the hydrogen peroxide group, with statistically significant differences ( t=20.69, 39.80, 12.73, all at P<0.05).Compared with the normal control group, the proportion of G2 phase cells in the hydrogen peroxide group significantly increased, showing a statistically significant difference ( t=23.10, P<0.05).The EDU-positive cell rate of hydrogen peroxide group was (25.41±6.99)%, significantly lower than (50.58±9.15)% of normal control group ( t=6.559, P<0.05).The relative expression level of lncRNA-p21 in the hydrogen peroxide group was 2.36±0.29, significantly higher than 1.02±0.02 in the normal control group ( t=7.893, P<0.05).The fluorescence in situ hybridization experiments indicate that lncRNA-p21 was localized in the cytoplasm. Conclusions:In the oxidative stress model induced by hydrogen peroxide, the proliferation ability of lens epithelial cells significantly decreases, the apoptosis level significantly increases, and the expression levels of ROS and lncRNA-p21 enhances.lncRNA-p21 may be involved in the oxidative stress injury process of lens epithelial cells.
