Expression of long-chain non-coding RNA-P21 in hydrogen peroxide induced human lens epithelial cells damage

Authors: Dong Xiaoming,  Liu Yuxuan,  Ji Liyang,  Wang Jing,  Zhang Jinsong
DOI: 10.3760/cma.j.cn115989-20221109-00522
Published 2024-03-10
Cite as Chin J Exp Ophthalmol, 2024, 42(3): 232-239.

Abstract                               [Download PDF] [Read Full Text]

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.

Key words:

Lens epithelial cells; Long-chain non-coding RNA-p21; Proliferation; Apoptosis; Oxidative stress

Contributor Information

Dong Xiaoming

Shenyang Aier Excellent Ophthalmology Hospital, Aier Group Ophthalmology Hospital Group Cataract and Artificial Lens Research Institute, Shenyang Aier Ophthalmology Precision Medical Research Institute, Ophthalmology Department of the Fourth Affiliated Hospital of China Medical University, China Medical University Ophthalmology Hospital, Liaoning Provincial Key Laboratory of Lens Research, Shenyang 110000, China

Liu Yuxuan

Shenyang Aier Excellent Ophthalmology Hospital, Aier Group Ophthalmology Hospital Group Cataract and Artificial Lens Research Institute, Shenyang Aier Ophthalmology Precision Medical Research Institute, Ophthalmology Department of the Fourth Affiliated Hospital of China Medical University, China Medical University Ophthalmology Hospital, Liaoning Provincial Key Laboratory of Lens Research, Shenyang 110000, China

Ji Liyang

Shenyang Aier Excellent Ophthalmology Hospital, Aier Group Ophthalmology Hospital Group Cataract and Artificial Lens Research Institute, Shenyang Aier Ophthalmology Precision Medical Research Institute, Ophthalmology Department of the Fourth Affiliated Hospital of China Medical University, China Medical University Ophthalmology Hospital, Liaoning Provincial Key Laboratory of Lens Research, Shenyang 110000, China

Wang Jing

Shenyang Aier Excellent Ophthalmology Hospital, Aier Group Ophthalmology Hospital Group Cataract and Artificial Lens Research Institute, Shenyang Aier Ophthalmology Precision Medical Research Institute, Ophthalmology Department of the Fourth Affiliated Hospital of China Medical University, China Medical University Ophthalmology Hospital, Liaoning Provincial Key Laboratory of Lens Research, Shenyang 110000, China

Aier College of Ophthalmology, Central South University, Changsha 410000, China

Zhang Jinsong

Shenyang Aier Excellent Ophthalmology Hospital, Aier Group Ophthalmology Hospital Group Cataract and Artificial Lens Research Institute, Shenyang Aier Ophthalmology Precision Medical Research Institute, Ophthalmology Department of the Fourth Affiliated Hospital of China Medical University, China Medical University Ophthalmology Hospital, Liaoning Provincial Key Laboratory of Lens Research, Shenyang 110000, China

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