Expression and effect of microRNA-2116-3p in human hypertrophic scar

doi:10.12307/2023.562

Abstract

Abstract: BACKGROUND: A wide range of studies have demonstrated that microRNA (miR) plays a crucial role in hypertrophic scars development, and type I collagen is closely related to hypertrophic scar fibroblasts. There might be a relationship between miR-2116-3p and occurrence and development of hypertrophic scars.
OBJECTIVE: To investigate the expression and role of miR-2116-3p on human hypertrophic scar fibroblasts.
METHODS: Hypertrophic scar tissues were collected from six patients from the First Affiliated Hospital of Xinjiang Medical University and normal skin tissues were collected from another six patients after blepharoplasty in the same unit at the same time. The expression of miR-2116-3p and type I collagen was detected by qRT-PCR. Hypertrophic scar tissues were taken and primary cells were cultured to passages 3 to 6. Hypertrophic scar fibroblasts were divided into negative control group, miR-2116-3p mimic group, and miR-2116-3p inhibitor group, and transfected with corresponding sequences. Cell proliferation was detected by cell counting kit-8 and EdU kit. Cell migration was detected by cell scratch assay, and apoptosis was detected by flow cytometry. Expression of type I collagen, type III collagen and α-smooth muscle actin was detected by western blot and qRT-PCR at protein and mRNA levels, respectively. Dual luciferase reporter assay system was used to carry out targeting assays.
RESULTS AND CONCLUSION: The mRNA expression of miR-2116-3p and type I collagen in hypertrophic scar tissues was significantly lower than that in normal skin tissues (P < 0.01). After 24, 48, and 72 hours of transfection, the cell viability of the miR-2116-3p mimic group was significantly lower than that of the negative control group (P < 0.05 or P < 0.01), while the cell viability of miR-2116-3p inhibitor group was significantly higher than that of the negative control group (P < 0.05 or P < 0.01). At 48 hours after transfection, the number of EdU positive cells in the miR-2116-3p mimic group was significantly lower than that in the negative control group, while the number of EdU positive cells in the miR-2116-3p inhibitor group was significantly higher than that in the negative control group (P < 0.05). At 24 hours after transfection, the cell scratch area and apoptotic rate of the miR-2116-3p mimic group were significantly higher than those of the negative control group (P < 0.01), and while the cell scratch area and apoptotic rate of the miR-2116-3p inhibitor group werer significantly lower than those of the negative control group (P < 0.01 or P < 0.05). At 24 hours after transfection, the mRMA and protein expression levels of type I collagen, type II collagen and α-smooth muscle actin were significantly decreased in the miR-2116-3p mimic group (P < 0.05 or P < 0.01) and significantly increased in the miR-2116-3p inhibitor group (P < 0.01) compared with the negative control group. Results from the dual luciferase assay showed that miR-2116-3p could bind to type I collagen. To conclude, the expression of miR-2116-3p is down-regulated in human hypertrophic scar. miR-2116-3p may inhibit the proliferation and migration of human hypertrophic scar fibroblasts and promote their apoptosis by targeting the expression of type I collagen.

Key words: miR-2116-3p, type I collagen, hypertrophic scar, fibroblast, skin fibrosis

CLC Number:

Tian Wenrong, Zuo Jun, Yu Yang, Qi Yusong, Ai Jiang, Bu Panpan, Zhao Jiaojun, Ma Zhiwei, Li Peipei, Ma Shaolin. Expression and effect of microRNA-2116-3p in human hypertrophic scar[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(28): 4480-4486.

Figures/Tables 7

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