miR-127-3p在动脉粥硬化中的表达及干扰miR-127-3p对PDGF-BB处理的HAoSMC增殖迁移的影响

Expression of miR-127-3p in atherosclerosis and the effect of interfering with miR-127-3p on proliferation and migration of PDGF-BB-treated HAoSMC

目的探讨miR-127-3p对血小板衍生生长因子-BB(PDGF-BB)诱导的人主动脉平滑肌细胞(HAoSMC)增殖、迁移的影响。方法收集动脉粥硬化患者及正常者血清,将HAoSMC分为对照组(未作任何处理)、模型组(25μg/L PDGF-BB)、anti-miR-NC组(转染miR-127-3p抑制剂对照+25μg/L PDGF-BB)、anti-miR-127-3p组(转染miR-127-3p抑制剂+25μg/L PDGF-BB)。实时荧光定量PCR(RT-qPCR)检测miR-127-3p和NF-κB mRNA表达水平;流式细胞仪检测细胞周期;四甲基偶氮唑盐比色法(MTT)检测细胞增殖活力;Transwell检测细胞迁移;蛋白质印迹(Western blot)法检测细胞增殖核抗原Ki67(Ki67)、神经钙粘蛋白(N-cadherin)、上皮钙粘蛋白(E-cadherin)的表达。结果与正常对照组相比,动脉粥硬化患者血清中miR-127-3p表达水平升高(P<0.05)。与对照组相比,模型组miR-127-3p表达水平升高,G0-G1期细胞所占比例降低,S期细胞所占比例升高,细胞OD值升高,迁移细胞数增加,Ki67、N-cadherin表达水平升高,E-cadherin表达水平降低(P<0.05)。与antimiR-NC组相比,anti-miR-127-3p组miR-127-3p表达水平降低,G0-G1期细胞所占比例升高,S期细胞所占比例降低,细胞OD值降低,迁移细胞数减少,Ki67、N-cadherin表达水平降低,E-cadherin表达水平升高,NF-κB mRNA表达水平降低(P<0.05)。结论干扰miR-127-3p表达可抑制PDGF-BB处理的HAoSMC增殖、迁移,其机制可能与NF-κB信号通路有关。

Objective To investigate the effect of miR-127-3p on platelet-derived growth factor-BB(PDGFBB)-induced proliferation and migration of human aortic smooth muscle cells(HAoSMC).Methods Serum was collected from atherosclerotic patients and normal subjects,and HAoSMC was divided into control group(without any treatment),model group(25μg/L PDGF-BB),anti-miR-NC group(control group transfected with miR-127-3p inhibitor+25μg/L PDGF-BB),anti-miR-127-3p group(transfected with miR-127-3p inhibitor+25μg/L PDGF-BB).Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression levels of miR-127-3p and NF-kappa B mRNA.Flow cytometry was used to detect cell cycle.Methyl thiazolyl tetrazolium(MTT)assay was used to detect cell proliferation activity.Transwell was used to detect cell migration.Western blot was used to detect the expression of proliferating nuclear antigen Ki67,N-cadherin and E-cadherin.Results Compared with the normal control group,the expression level of serum miR-127-3p was increased in atherosclerotic patients(P<0.05).Compared with the control group,the expression level of miR-127-3p increased,the proportion of cells in G0-G1 phase decreased,the proportion of cells in S phase increased,the OD value of cells increased,the number of migrating cells increased,the expression levels of Ki67,N-cadherin increased,and the expression level of E-cadherin decreased in the model group(P<0.05).Compared with the anti-miR-NC group,the anti-miR-127-3p group had lower expression level of miR-127-3p,higher proportion of cells in G0-G1 phase,lower proportion of cells in S phase,lower OD value of cells,lower number of migrating cells,lower expression level of Ki67,N-cadherin,higher expression level of E-cadherin and lower expression level of NF-kappa B mRNA(P<0.05).Conclusion Interference with the expression of miR-127-3p can inhibit the proliferation and migration of PDGF-BB-treated HAoSMC,and the mechanism may be related to the NF-kappa B signaling pathway.