摘要
目的:探讨环状RNA(circular RNA,circRNA)hsa_circ_0013958对人瘢痕疙瘩成纤维细胞(Human keloid fibroblast,HKF)增殖、迁移、侵袭的影响及分子机制.方法:将人瘢痕疙瘩成纤维细胞分为si-NC组、si-hsa_circ_0013958组、miR-NC组、miR-637组、si-hsa_circ_0013958+anti-miR-NC组和si-hsa_circ_0013958+anti-miR-637组.实时荧光定量PCR(Real time quantitative polymerase chain reaction,RT-qPCR)试剂盒检测hsa_circ_0013958、miR-637表达;四甲基偶氮唑盐(Methyl thiazolyl tetrazolium,MTT)比色法检测细胞存活率;平板克隆实验检测细胞集落形成数;Transwell检测细胞迁移和侵袭能力;双荧光素酶报告实验检测hsa_circ_0013958和miR-637的靶向关系.结果:瘢痕疙瘩组织中hsa_circ_0013958表达水平较正常皮肤组织升高,miR-637表达水平降低(P＜0.05).干扰hsa_circ_0013958表达或过表达miR-637后,HKF细胞存活率、集落形成数以及迁移、侵袭细胞数均降低(P＜0.05).hsa_circ_0013958和miR-637有靶向调控关系;抑制miR-637表达逆转了干扰hsa_circ_0013958对HKF增殖、迁移侵袭的影响.结论:干扰hsa_circ_0013958通过调控miR-637抑制人瘢痕疙瘩成纤维细胞的增殖、迁移和侵袭.
Abstract
Objective To explore the effect of circular RNA(circRNA)hsa_circ_0013958 on the proliferation,migration and invasion of human keloid fibroblasts and its molecular mechanism.Methods Human keloid fibroblasts HKF were divided into si-NC group,si-hsa_circ_0013958 group,miR-NC group,miR-637 group,si-hsa_circ_0013958+anti-miR-NC group,si-hsa_circ_0013958+anti-miR-637 group.Expression levels of hsa_circ_0013958 and miR-637 were detected by Real time quantitative polymerase chain reaction(RT-qPCR).Cell viability was assayed by Methyl thiazolyl tetrazolium(MTT)method,cell colony formation was detected by plate cloning test,cell migration and invasion ability was detected by transwell method,the targeting relationship between hsa_circ_0013958 and miR-637 was detected by dual luciferase reporter experiment.Results Compared with normal skin tissues,the expression level of hsa_circ_0013958 were elevated in keloid tissue,and the expression of miR-637 was decreased(P＜0.05).After interference with hsa_circ_0013958 expression or overexpression of miR-637,the survival rate,colony forming number,migration and invasion of HKF cells were decreased(P＜0.05).Hsa_circ_0013958 has a targeted regulatory relationship with miR-637,inhibition of miR-637 expression reversed the effect of interference with hsa_circ_0013958 on the proliferation,migration and invasion of HKF.Conclusion Interference with hsa_circ_0013958 may inhibit the proliferation,migration and invasion of human keloid fibroblasts by regulating miR-637.
国家科技期刊平台
NSTL
维普
万方数据