microRNA-132对人脂肪源性间充质干细胞成骨分化的影响

Effect of microRNA-132 on the osteogenic differentiation of human adipose derived mesenchymal stem cells

目的:探讨microRNA-132(miR-132)对人脂肪源性间充质干细胞(hADSCs)成骨分化的影响。方法:分离培养hAD-SCs,转染过表达或抑制miR-132(miR-132mimics或miR-132inhibitors),分别采用qPCR法和western blotting法检测细胞中成骨相关基因碱性磷酸酶(ALP)、骨钙素(OCN)、Runt相关转录因子2(Runx2)、Osterix mRNA和蛋白相对表达量。结果:miR-132在hADSCs的表达随着成骨诱导时间延长逐渐升高(P<0.05),转染miR-132 mimics后,hADSCs成骨相关基因ALP、OCN、Runx2、Osterix mRNA和蛋白的表达显著升高,而转染miR-132inhibitors后,ALP、OCN、Runx2、Osterix mRNA和蛋白的表达显著降低,差异均有统计学意义(均P<0.05)。结论:miR-132通过正向调控hADSCs成骨相关基因ALP、OCN、Runx2、Osterix表达进而影响hADSCs的成骨分化进程。

Objective:To explore the effect of microRNA-132(miR-132) on the osteogenic differentiation of human adipose derived mesenchymal stem cells(hADSCs). Methods:The hADSCs were isolated and cultured.Then the cells were transfected with mimic negative control,miR-132 mimics,an inhibitor negative control,or miR-132 inhibitors.The protein and mRNA expressions of alkaline phosphatase(ALP),osteocalcin(OCN),Runt related transcription factor 2(Runx2) and Osterix were detected by qPCR and western blotting,respectively. Results:The expression of miR-132 increased gradually with the prolongation of osteogenic induction time( P <0.05).Overexpression of miR-132 resulted in up-regulation of ALP,OCN,Runx2 and Osterix in hADSCs,whereas inhibition of miR-132 attenuated the expressions of ALP,OCN,Runx2 and Osterix( P <0.05). Conclusion:miR-132 might promote the osteogenic differentiation of hADSCs via up-regulation of osteoblast-related genes (ALP,OCN,Runx2 and Osterix).