Objective To explore the effect of long non-coding RNA small nucleolar RNA host gene 1 (lncRNA SNHG1) on the proliferation of human hypertrophic scar fibroblasts. Methods Twenty-two patients with hypertrophic scars who underwent surgical treatment in the Department of Burns and Plastic Surgery of the General Hospital of Ningxia Medical University from 2019 to 2021 were included. The hypertrophic scar tissues and the normal skin tissues adjacent to the scars (within 3 cm adjacent to the scars) were collected, and fibroblasts were isolated and cultured from them. The relative expression levels of SNHG1 mRNA and miR - 382 - 3p were detected by qRT - PCR at the tissue and cell levels. The hypertrophic scar fibroblasts were randomly divided into a control group, an SNHG1 negative control group (transfected with a lentiviral empty vector), a mimic control group (transfected with a control mimic), an SNHG1 negative control + mimic control group (transfected with a lentiviral empty vector and a control mimic), an SNHG1 overexpression group (transfected with an overexpression lentivirus), a miR - 382 - 3p overexpression group (transfected with a miR - 382 - 3p mimic), an SNHG1 overexpression + mimic control group (transfected with an overexpression lentivirus and a control mimic), and an SNHG1 overexpression + miR - 382 - 3p overexpression group (transfected with an overexpression lentivirus and a miR - 382 - 3p mimic). The relative expression levels of SNHG1, PCNA, p27 mRNA and miR - 382 - 3p in each group were detected by qRT - PCR; the cell proliferation ability in each group was detected by the CCK - 8 method; the cell proliferation level in each group was detected by the EdU staining method; the protein expression levels of p27 and PCNA in each group were detected by Western blotting. Results Compared with normal skin tissues and their fibroblasts, SNHG1 mRNA was highly expressed in hypertrophic scar tissues (3.21 ± 2.65 vs. 1.14 ± 0.61, P < 0.001) and their fibroblasts (0.91 ± 0.08 vs. 0.54 ± 0.08, P < 0.01), while the expression of miR - 382 - 3p was downregulated (tissue: 0.53 ± 0.34 vs. 1.15 ± 0.61, P < 0.001; cell: 0.84 ± 0.09 vs. 1.01 ± 0.004, P < 0.05). Compared with the SNHG1 negative control group, the cell proliferation ability in the SNHG1 overexpression group was enhanced (0.23 ± 0.03 vs. 0.16 ± 0.01, P < 0.001), the percentage of EdU-positive cells was significantly increased (30.01% ± 5.70% vs. 7.13% ± 4.40%, P < 0.001), the relative expression levels of PCNA mRNA and protein were increased (mRNA: 2.97 ± 0.33 vs. 0.98 ± 0.25, P < 0.01; protein: 2.20 ± 0.09 vs. 0.88 ± 0.20, P < 0.05), the relative expression levels of p27 mRNA and protein were decreased (mRNA: 0.30 ± 0.03 vs. 1.42 ± 0.15, P < 0.001; protein: 0.47 ± 0.11 vs. 1.13 ± 0.19, P < 0.05), and the relative expression level of miR - 382 - 3p was decreased (0.05 ± 0.01 vs. 1.03 ± 0.12, P < 0.001); compared with the SNHG1 overexpression + mimic control group, the cell proliferation ability in the SNHG1 overexpression + miR - 382 - 3p overexpression group was weakened (0.15 ± 0.02 vs. 0.26 ± 0.01, P < 0.001), the percentage of EdU-positive cells was decreased (5.97% ± 0.33% vs. 11.70% ± 0.87%, P < 0.001), the relative expression levels of PCNA mRNA and protein were decreased (mRNA: 0.64 ± 0.09 vs. 3.33 ± 0.38, P < 0.001; protein: 1.70 ± 0.36 vs. 2.34 ± 0.16, P < 0.05), and the relative expression levels of p27 mRNA and protein were increased (mRNA: 1.01 ± 0.44 vs. 0.09 ± 0.04, P < 0.05; protein: 1.38 ± 0.31 vs. 0.50 ± 0.09, P < 0.05). Conclusion SNHG1 is highly expressed in hypertrophic scars and can negatively regulate the expression of miR - 382 - 3p, thereby promoting the proliferation of hypertrophic scar fibroblasts and is expected to become a new target for the treatment of hypertrophic scars.
目的探讨长链非编码RNA核仁小RNA宿主基因1(lncRNA SNHG1)对人增生性瘢痕成纤维细胞增殖的影响。方法纳入2019-2021年在宁夏医科大学总医院烧伤整形外科接受手术治疗的22例增生性瘢痕患者,收集其增生性瘢痕组织与瘢痕旁正常皮肤组织(瘢痕旁3 cm以内),从中分离培养成纤维细胞。采用qRT-PCR从组织和细胞水平检测SNHG1 mRNA和miR-382-3p相对表达水平。将增生性瘢痕成纤维细胞随机分为对照组、SNHG1阴性对照组(转染慢病毒空载体)、模拟物对照组(转染对照模拟物)、SNHG1阴性对照+模拟物对照组(转染慢病毒空载体和对照模拟物)、SNHG1过表达组(转染过表达慢病毒)、miR-382-3p过表达组(转染miR-382-3p模拟物)、SNHG1过表达+模拟物对照组(转染过表达慢病毒和对照模拟物)与SNHG1过表达+miR-382-3p过表达组(转染过表达慢病毒和miR-382-3p模拟物),采用qRT-PCR检测各组SNHG1、PCNA、p27 mRNA和miR-382-3p的相对表达水平;CCK-8法检测各组细胞增殖能力;EdU染色法检测各组细胞增殖水平;Western blotting检测各组p27和PCNA蛋白表达水平。结果与正常皮肤组织及其成纤维细胞相比,SNHG1 mRNA在增生性瘢痕组织(3.21±2.65 vs.1.14±0.61,P<0.001)及其成纤维细胞中呈高表达(0.91±0.08 vs.0.54±0.08,P<0.01),而miR-382-3p表达下调(组织:0.53±0.34 vs.1.15±0.61,P<0.001;细胞:0.84±0.09 vs.1.01±0.004,P<0.05)。与SNHG1阴性对照组相比,SNHG1过表达组细胞增殖能力增强(0.23±0.03 vs.0.16±0.01,P<0.001),EdU阳性细胞百分比明显增高(30.01%±5.70% vs.7.13%±4.40%,P<0.001),PCNA mRNA和蛋白相对表达水平增高(mRNA:2.97±0.33 vs.0.98±0.25,P<0.01;蛋白:2.20±0.09 vs.0.88±0.20,P<0.05),p27 mRNA和蛋白相对表达水平降低(mRNA:0.30±0.03 vs.1.42±0.15,P<0.001;蛋白:0.47±0.11 vs.1.13±0.19,P<0.05),miR-382-3p相对表达水平降低(0.05±0.01 vs.1.03±0.12,P<0.001);与SNHG1过表达+模拟物对照组相比,SNHG1过表达+miR-382-3p过表达组细胞增殖能力减弱(0.15±0.02 vs.0.26±0.01,P<0.001),EdU阳性细胞百分比下降(5.97%±0.33% vs.11.70%±0.87%,P<0.001),PCNA mRNA和蛋白相对表达水平降低(mRNA:0.64±0.09 vs.3.33±0.38,P<0.001;蛋白:1.70±0.36 vs.2.34±0.16,P<0.05),p27 mRNA和蛋白相对表达水平升高(mRNA:1.01±0.44 vs.0.09±0.04,P<0.05;蛋白:1.38±0.31 vs.0.50±0.09,P<0.05)。结论SNHG1在增生性瘢痕中呈高表达,可负向调控miR-382-3p的表达,进而促进增生性瘢痕成纤维细胞的增殖,有望成为治疗增生性瘢痕的新靶点。
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