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目的:探讨microRNA-382(miR-382)对脐带间充质干细胞(hUC-MSC)生物学特性的影响,以更深入的了解miRNA在MSC生物学特性中的调节机制。方法:对贴壁培养的hUC-MSC分别以miR-382 mimics及miR-382 inhibitor转染,在倒置显微镜下观察细胞形态变化;使用CCK-8检测miR-382转染对hUC-MSC增殖的影响;分别使用油红O和茜素红染色检测miR-382对hUC-MSC成脂及成骨分化的影响,并使用氯化十六烷基吡啶溶解茜素红检测其吸光度值以定量分析成骨情况,以RT-PCR检测Runx2基因表达情况;使用RT-PCR方法检测miR-382对hUCMSC分泌细胞因子基因表达的影响。结果:miR-382对hUC-MSC的形态无影响;miR-382不影响hUC-MSC的增殖;miR-382可以抑制hUC-MSC成骨分化而不影响其成脂分化;miR-382可以影响hUC-MSC细胞因子基因的表达,如IL-6,IDO1,G-CSF,M-CSF和GM-CSF的表达。结论:miR-382可以抑制hUC-MSC的成骨分化,并对其所分泌细胞因子的基因表达产生影响。
OBJECTIVE: To investigate the effect of microRNA-382 (miR-382) on the biological characteristics of umbilical cord mesenchymal stem cells (hUC-MSCs) to further understand the regulatory mechanism of miRNAs in the biological characteristics of MSC. Methods: Adherent cells were transfected with miR-382 mimics and miR-382 inhibitor respectively. The morphological changes of cells were observed under an inverted microscope. The effect of miR-382 transfection on the proliferation of hUC-MSCs was detected by CCK-8 The effects of miR-382 on adipogenic and osteogenic differentiation of hUC-MSCs were detected by oil red O and alizarin red staining, respectively. The absorbance values of alizarin red were determined by cetylpyridinium chloride dissolved to quantitatively analyze the osteogenic The expression of Runx2 gene was detected by RT-PCR. The effect of miR-382 on the gene expression of cytokine secreted by hUCMSC was detected by RT-PCR. Results: miR-382 had no effect on the morphology of hUC-MSCs; miR-382 did not affect the proliferation of hUC-MSCs; miR-382 inhibited osteogenic differentiation of hUC-MSCs without affecting adipogenic differentiation; miR-382 could affect hUC Expression of MSC cytokine genes such as IL-6, IDO1, G-CSF, M-CSF and GM-CSF. Conclusion: miR-382 can inhibit the osteogenic differentiation of hUC-MSC and affect the gene expression of cytokines secreted by hUC-382.