干扰SNHG7对人瘢痕疙瘩成纤维细胞增殖和凋亡的影响  

Effects of interfering with SNHG7 on proliferation and apoptosis of human keloid fibroblasts

作　　者：任永强[1,2] 李庆华[1] 黄新建[1] REN Yong-Qiang;LI Qing-Hua;HUANG Xin-Jian(School of Clinical Medicine,Henan University of Science and Technology,Luoyang 471003,China)

机构地区：[1]河南科技大学临床医学院,洛阳471003 [2]河南科技大学第一附属医院整形美容科,洛阳471003

出　　处：《中国免疫学杂志》2020年第15期1819-1824,共6页Chinese Journal of Immunology

摘　　要：目的:探究干扰小核仁RNA宿主基因7(SNHG7)对人瘢痕疙瘩成纤维细胞增殖和凋亡的影响及作用机制。方法:分离培养人瘢痕疙瘩成纤维(KF)细胞,将si-NC、si-SNHG7、pcDNA3.1、pcDNA3.1-SNHG7、miR-NC、miR-122分别转染至KF细胞,记为si-NC组(SNHG7阴性对照组)、si-SNHG7组(SNHG7抑制表达组)、pcDNA3.1组(SNHG7阳性对照组)、pcDNA3.1-SNHG7组(SNHG7过表达组)、miR-NC组(miR-122阳性对照组)、miR-122组(miR-122过表达组)。将si-SNHG7分别与anti-miR-NC、anti-miR-122共转染至KF细胞中,记为si-SNHG7+anti-miR-NC组(miR-122阴性对照组)、si-SNHG7+anti-miR-122组(miR-122抑制剂组)。RT-qPCR检测SNHG7和miR-122表达水平;Western blot检测细胞周期素(CyclinD1)、细胞周期蛋白依赖性激酶抑制剂1A(P21)、Bcl-2相关X蛋白(Bax)、B细胞淋巴瘤/白血病-2(Bcl-2)蛋白表达水平;MTT检测细胞增殖抑制率;流式细胞术检测细胞凋亡;双荧光素酶报告实验检测SNHG7和miR-122的靶向关系。结果:与正常皮肤组织相比,瘢痕疙瘩组织SNHG7表达显著升高,miR-122表达显著降低(P<0.05)。抑制SNHG7表达和过表达miR-122,CyclinD1、Bcl-2表达显著降低,P21、Bax表达显著升高,细胞增殖抑制率显著升高,细胞凋亡率显著升高(P<0.05)。SNHG7靶向调控miR-122,抑制miR-122表达逆转SNHG7对KF增殖和凋亡的作用。结论:抑制SNHG7表达可抑制KF细胞增殖,促进其凋亡,具体机制可能与miR-122有关,为瘢痕疙瘩的治疗提供新思路和新靶点。Objective:To investigate effect and mechanism of interfering with small nucleolar RNA host gene 7(SNHG7)on proliferation and apoptosis of human keloid fibroblasts.Methods:Human keloid fibroblasts(KF)cells were isolated and cultured,and si-NC,si-SNHG7,pcDNA3.1,pcDNA3.1-SNHG7,miR-NC,and miR-122 were transfected into KF cells,respectively,which were recorded as si-NC group(SNHG7 negative control group),si-SNHG7 group(SNHG7 inhibition expression group),pcDNA3.1 group(SNHG7 positive control group),pcDNA3.1-SNHG7 group(SNHG7 overexpression group),miR-NC group(miR-122 positive control group),miR-122 group(miR-122 overexpression group),and si-SNHG7 was co-transfected into KF cells with anti-miR-NC and anti-miR-122,respectively,recorded as si-SNHG7+anti-miR-NC group(miR-122 negative control group)and si-SNHG7+anti-miR-122 group(miR-122 inhibitor group).RT-qPCR was used to detect expressions of SNHG7 and miR-122;Western blot was used to detect CyclinD1 and cyclin-dependent kinase inhibitor 1A(P21),Bcl-2 assiociated X protein(Bax),B cell lymphoma/leukemia-2(Bcl-2)protein expressions;MTT investigated cell proliferation inhibition rate;Flow cytometry detected apoptosis;Dual luciferase reporter assay was used to observe targeting relationship between SNHG7 and miR-122.Results:Compared with normal skin tissue,expression of SNHG7 in keloid tissue was significantly increased,and expression of miR-122 was significantly decreased(P<0.05).Inhibition of SNHG7 expression and overexpression of miR-122,CyclinD1 and Bcl-2 expressions were significantly decreased,P21 and Bax expressions were significantly increased,cell proliferation inhibition rate was obviously increased,and apoptosis rate was obviously increased(P<0.05).SNHG7 targeting miR-122 and inhibiting expression of miR-122 reversed effect of SNHG7 on KF cells proliferation and apoptosis.Conclusion:Inhibition of SNHG7 expression can inhibit proliferation of and promote cell apoptosis,whose mechanism may be related to miR-122,which will provide new ideas and targets for treatment

关 键 词：SNHG7 MIR-122 瘢痕疙瘩成纤维细胞 增殖 凋亡 

分 类 号：R619.6[医药卫生—外科学]