MicroRNA-2116-3p Regulates The Viability Of Fibroblasts In Hypertrophic Scars

Objective:To investigate the expression and function of miR-2116-3p in human hypertrophic scars.Methods:Hypertrophic scar tissues from 6 eligible patients who visited the First Affiliated Hospital of Xinjiang Medical University and normal skin tissues removed after eyelid surgery from 6 eligible patients who visited the same hospital at the same time were collected.Real-time fluorescent quantitative PCR was used to detect the expression of miR-2116-3p and Col1.Hypertrophic scar fibroblasts were primary cultured from hypertrophic scar tissues and passaged to P₃-P₆for experiments.The cells were divided into a negative control group,a miR-2116-3p mimic group,and a miR-2116-3p inhibitor group,respectively.The corresponding sequences were transfected and the cell proliferation activity was detected by CCK-8 assay and EDU staining.The cell migration ability was detected by scratch assay.Cell apoptosis was detected by flow cytometry.The gene and protein expression levels of Col1,Col3,andα-smooth muscle actin were detected by real-time fluorescent quantitative PCR and Western blot,respectively.The dual-luciferase experiment was used to verify the target binding.Results:（1）The m RNA expression of miR-2116-3p was significantly lower in hypertrophic scar tissues than in normal skin tissues,while the m RNA expression of Col1 was significantly higher in hypertrophic scar tissues than in normal skin tissues.（2）After 48 hours of transfection,the cell viability in the mimic group was significantly lower than that in the negative control group,and the cell viability in the inhibitor group was significantly higher than that in the negative control group.（3）After 48 hours of transfection,the number of EDU-labeled cells in the mimic group was significantly lower than that in the negative control group,and the number of EDU-labeled cells in the inhibitor group was significantly higher than that in the negative control group.（4）After 24 hours of transfection,the scratch area of cells in the mimic group was significantly larger than that in the negative control group,and the scratch area of cells in the inhibitor group was significantly smaller than that in the negative control group.（5）After 48 hours of transfection,the apoptosis rate in the mimic group significantly increased,while the apoptosis rate in the inhibitor group significantly decreased.（6）After 48 hours of transfection,the gene and protein expression of Col1,Col3,andα-SMA in the mimic group significantly decreased,while that in the inhibitor group significantly increased.（7）The dual-luciferase experiment showed that miR-2116-3p could target binding to Col1.Conclusion:The expression of miR-2116-3p is downregulated in human hypertrophic scars.miR-2116-3p may inhibit the proliferation and migration of human hypertrophic scar fibroblasts and promote their apoptosis by targeting Col1.This provides a theoretical basis for finding effective targets for the treatment of hypertrophic scars in the future.