敲低环状RNA WD重复含蛋白1抑制膝骨关节炎软骨细胞增殖并诱导凋亡

Knockdown of circRNA WD repeat containing protein 1 inhibits proliferation and induces apoptosis of chondrocytes in knee osteoarthritis

二维码有效期 120s

摘要背景:环状RNA(circular RNA,circRNA)在多种疾病或肿瘤中发挥着重要作用,近年研究结果发现,膝骨关节炎中circRNA失常表达,并可通过调控微小RNA(miRNA)/mRNA参与疾病进展.目的:探究circ-BRWD1/miR-488-3p/DNMT3A对膝骨关节炎软骨细胞增殖和凋亡的影响.方法:实时荧光定量PCR检测膝骨关节炎软骨细胞miR-488-3p、circ-BRWD1、DNMT3A表达.将细胞分为si-NC组、si-circ-BRWD1组、si-circ-BRWD1+ anti-miR-NC组、si-circ-BRWD1+anti-miR-488-3p组、vector组、circ-BRWD1组、miR-NC组、miR-488-3p组、miR-488-3p+vector组、miR-488-3p+ DNMT3A组、anti-miR-NC组、anti-miR-488-3p组;实时荧光定量PCR检测circ-BRWD1、miR-488-3p、DNMT3A表达,MTT、流式细胞术检测增殖和凋亡,Western blot检测DNMT3A、增殖、凋亡相关蛋白表达;双荧光素酶报告实验检测circ-BRWD1与miR-488-3p、miR-488-3p与DNMT3A的靶向关系.结果与结论:①与正常软骨细胞相比,circ-BRWD1、DNMT3A在膝骨关节炎软骨细胞中高表达,miR-488-3p低表达;②敲低circ-BRWD1或过表达miR-488-3p可抑制膝骨关节炎软骨细胞增殖并诱导凋亡;③circ-BRWD1靶向负调控miR-488-3p,抑制miR-488-3p可逆转敲低circ-BRWD1对膝骨关节炎软骨细胞增殖、凋亡的影响;④miR-488-3p靶向负调控DNMT3A,上调DNMT3A可逆转过表达miR-488-3p对膝骨关节炎软骨细胞增殖、凋亡的影响;⑤circ-BRWD1可通过调控miR-488-3p进而调控DNMT3A的表达;⑥结果表明,敲低circ-BRWD1可抑制膝骨关节炎软骨细胞增殖并诱导凋亡,其作用机制可能与调控miR-488-3p/DNMT3A轴有关.