Effect and Regulatory Mechanism of CircHIPK3/miR-330-5p on Injury of Human Lens Epithelial Cells
Objective To explore the effect and regulatory mechanisms of circHIPK3 and miR-330-5p expression on human lens epithelial cells injury.Methods Human lens epithelial cells(LECs)SRA01/04 were used as the cell experimental material,and a cell injury model of SRA01/04 was induced with hydrogen peroxide(H2O2).The SRA01/04 cells were randomly divided into 6 groups:NC group,model group,overexpression plasmid(OE-Vector)+H2O2 group,circHIPK3 overexpression plasmid(OE-circHIPK3)+H2O2 group,OE-circHIPK3+miR-330-5p negative control mimic NC sequence(miR-NC)+H2O2 group and OE-circHIPK3+miR-330-5p oligonucleotide mimics(miR-330-5p mimics)+H2O2 group.The qRT-PCR method was used to detect the levels of circHIPK3 and miR-330-5p in each group.The dual luciferase reporter assay was used to verify the targeting relationship between circHIPK3 and miR-330-5p.CCK-8 method and flow cytometry were used to detect cell survival rate and apoptosis rate respectively.The levels of superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione peroxidase(GSH-Px)were detected.Western blot was used to detect the expression of B lymphomatoma-2 related protein(Bax)and B lymphomatoma-2(Bcl-2).Results The level of circHIPK3 was lower and the level of miR-330-5p was higher in the model group cells than in the NC group(P<0.05).Compared with those in the NC group,the cell survival rate,Bcl-2 protein level,SOD and GSH-Px level decreased in the model group,but apoptosis rate,Bax protein level and MDA level increased(P<0.05).Compared with those in the model group and OE-Vector+H2O2 group,the cell survival rate in the OE-circHIPK3+H2O2 group increased,the apoptosis rate and Bax protein level decreased,the Bcl-2 protein level climbed,and the SOD,GSH-Px and MDA levels dropped(P<0.05).CircHIPK3 was found with the ability to target the expression of miR-330-5p.Compared with those in the OE-circHIPK3+miR-NC+H2O2 group,the miR-330-5p level of the OE-circHIPK3+miR-330-5p mimics+H2O2 group climbed,but the cell survival rate,Bcl-2 protein level,SOD and GSH-Px levels dropped.Moreover,the apoptosis rate and levels of Bax protein and MDA increased(P<0.05).Conclusions Overexpression of circHIPK3 will target the inhibition of miR-330-5p expression,promote LECs survival,inhibit LECs apoptosis,enhance their antioxidant stress resistance and alleviate LECs damage.
Human lens epithelial cellsCircular RNACircular RNA homologous domain interacting protein kinase 3Mi-cro RNA-330-5p