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| MiR-21 Synergizes with BMP9 in Osteogenic Differentiation During Mesenchymal Stem Cells Line C3H10T1/2 |
| SONG Qi-ling, SHI Qiong, CHEN Chu, TANG Zu-chuan, LIU Hong-xia, ZHOU Yi-qing, ZHANG Ru-yi, YAN Shu-juan, WENG Ya-guang |
| Key Laboratory of Clinical Laboratory Diagnostics of Ministry Education, Faculty of Laboratory Medicine, Chongqing Medicine University, Chongqing 400016, China |
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Abstract Objective:(1) To study if there is a crosstalk between bone morphogenetic protein 9(BMP9) and microRNA-21(miR-21);(2) to investigate the effect of miR-21 on BMP9-induced osteogenic differentiation of mesenchymal stem cells line C3H10T1/2.Methods:(1) Using Ad-BMP9 infecting C3H10 T1/2 cells, the expression of miR-21 were detected by Real-time-PCR. After transfecting miR-21 mimic 1 day and 5 days, the expression of BMP were tested by Real-time-PCR.(2) In vitro, after transfecting miR-21, BMP9-CM treating C3H10T1/2, ALP staining and ALP activity were detected and the calcium deposition were detected by Alizarin Red S staining.(3)The expression of BMP9 downstream factors ALP and OCN were detected by Real-time-PCR.(4) The protein expression of p-Smad1/5 were detected by Western blot. Results:BMP9 can decrease the expression of miR-21 temporarily and miR-21 also decreases the expression of BMP9 temporarily. After transfecting miR-21 and treating C3H10T1/2 cell with BMP9-CM, the early osteogenic differentiation marker ALP activity and ALP staining and later osteogenic differentiation marker calcium deposition were enhanced. What's more, the expression of ostengenic factors ALP and OCN also increase during miR-21 combined with BMP9-CM. The protein expression of p-Smad1/5 were upregulation during the same process. Conclusion:There is a crosswalk between BMP9 and miR-21. MiR-21 synergizes with BMP9 promoting mesenchymal stem cells C3H10T1/2 osteogenic differentiation by increasing BMP9/Smad signaling pathway activated.
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Received: 27 July 2015
Published: 19 November 2015
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Cite this article:
SONG Qi-ling, SHI Qiong, CHEN Chu, TANG Zu-chuan, LIU Hong-xia, ZHOU Yi-qing, ZHANG Ru-yi, YAN Shu-juan, WENG Ya-guang. MiR-21 Synergizes with BMP9 in Osteogenic Differentiation During Mesenchymal Stem Cells Line C3H10T1/2. China Biotechnology, 2016, 36(2): 22-29.
URL:
https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20160204 OR https://manu60.magtech.com.cn/biotech/Y2016/V36/I2/22
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