Inhibitory effect of hsa-miR-199a-5p mediated by lentivirus on keloid fibroblast fibrosis through TGF-β/Smad pathway
10.3760/cma.j.cn114453-20210422-00179
- VernacularTitle:慢病毒介导hsa-miR-199a-5p通过转化生长因子-β/Smad通路抑制瘢痕疙瘩成纤维细胞纤维化的研究
- Author:
Yucang SHI
1
;
Zhihong ZHOU
;
Yanfang LIU
;
Zeyong WU
;
Haihua HUANG
;
Zhiyuan WU
Author Information
1. 广东医科大学附属医院整形外科，湛江 524001
- Keywords:
Keloid;
Fibroblasts;
MicroRNAs;
hsa-miR-199a-5p;
Collagen;
Fibrosis
- From:
Chinese Journal of Plastic Surgery
2022;38(5):570-577
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of hsa-miR-199a-5p on keloid fibroblasts (KFs) fibrosis and its relationship with TGF-β/Smad signaling pathway in vitro. Methods:KFs were infected with the lentiviral vector carrying hsa-miR-199a-5p (hsa-miR-199a-5p group) or the blank vector (rLv-NC group), and a blank control group was set. The proliferation status of KFs were detected by CCK-8 assay. At 48 hours after infection, the expression of hsa-miR-199a-5p in KFs were detected by quantitative real-time PCR (qRT-PCR); cell apoptosis was detected by flow cytometry analysis; cell invasion ability was detected by Transwell; The mRNA and protein levels of collagen Ⅰ, collagen Ⅲ, α-smooth muscle actin(α-SMA), Smad3 and TGF-β1 were detected by qRT-PCR and Western blotting, respectively.Results:At 48 hours after infection with RLV-hsa-miR-199a-5p lentiviral vector, the mRNA expression level of hsa-miR-199a-5p in hsa-miR-199a-5p group KFs was significantly higher than that in rLv-NC group and blank control group, and the difference was statistically significant ( P<0.01). Compared with the rLv-NC and blank control groups, the proliferation rate and invasiveness of the hsa-miR-199a-5p group decreased, the apoptosis rate increased, with statistical significance ( P<0.01); the mRNA and protein expression levels of collagen Ⅰ, collagen Ⅲ, α-SMA, Smad3 and TGF-β1 in hsa-miR-199a-5p group were significantly lower than those in rLv-NC group and blank control group, with statistical significance ( P<0.01). Conclusions:hsa-miR-199a-5p may inhibit the proliferation and invasion of KFs by inhibiting TGF-β/Smad pathway, and promote the apoptosis of KFs, thus inhibiting keloid fibrosis.
