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Effect of miR-181a-3p on osteogenic differentiation of human bone marrow-derived mesenchymal stem cells by targeting BMP10

机译:miR-181a-3p靶向BMP10对人骨髓间充质干细胞成骨分化的影响

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Objective: To explore the regulation relationship between miR-181a-3p and BMP10, and their mechanism of osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (MSCs). Methods: After osteogenic induction of MSCs, the ALP activity was detected by ELISA. The expression of miRNA-181a-3p and BMP10 was detected by RT-qPCR, and the protein levels of BMP10 and osteogenic differentiation marker proteins ALK and RUNX2 were detected by Western blot. The TargetScan online website was used to predict the putative target of miR-181a-3p, and dual luciferase reporter assay was performed to validate the targeting relationship between miR-181a-3p and BMP10. Results: In osteogenic differentiation of MSCs, ALP activity, the level of ALK and RUNX2 was evidently increased ( p ?.05), and the expression of miR-181a-3p was significantly downregulated ( p ?.05). Moreover, overexpression of miR-181a-3p obviously decreased the expression of BMP10 ( p ?.05), miR-181a-3p knockdown increased the expression of BMP10 prominently ( p ?.05). The transfection of miR-181a-3p mimics resulted in significantly downregulation of ALP activity and RUNX2 protein expression in MSCs ( p ?.05). In addition, overexpression of BMP10 could reverse the inhibitory effect of miR-181a-3p on osteogenic differentiation ( p ?.05). Conclusions: In conclusion, we found that miR-181a-3p inhibited osteogenic differentiation of MCSs by targeting BMP10.
机译:目的:探讨miR-181a-3p和BMP10之间的调控关系,以及其与人骨髓间充质干细胞(MSCs)成骨分化的机制。方法:成骨诱导骨髓间充质干细胞后,ELISA法检测其ALP活性。 RT-qPCR检测miRNA-181a-3p和BMP10的表达,Western blot检测BMP10,成骨分化标记蛋白ALK和RUNX2的表达。使用TargetScan在线网站预测miR-181a-3p的假定靶标,并进行双荧光素酶报告基因分析以验证miR-181a-3p与BMP10之间的靶向关系。结果:在MSC的成骨分化中,ALP活性,ALK和RUNX2的水平明显升高(p <0.05),而miR-181a-3p的表达显着下调(p <0.05)。而且,miR-181a-3p的过表达明显降低了BMP10的表达(p <?0.05),miR-181a-3p的敲低显着提高了BMP10的表达(p <?0.05)。 miR-181a-3p模拟物的转染导致MSC中ALP活性和RUNX2蛋白表达显着下调(p <?0.05)。此外,BMP10的过表达可以逆转miR-181a-3p对成骨分化的抑制作用(p <?0.05)。结论:总之,我们发现miR-181a-3p通过靶向BMP10抑制MCS的成骨分化。

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