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Hyperin up-regulates miR-7031-5P to promote osteogenic differentiation of MC3T3-E1 cells
Dongchen Qian1,2, Yueyue Chen3, Xusheng Qiu4
1Department of Orthopedic, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing, PR China.
Objective:
To investigate the effects of Hyperin (Hyp) on osteogenic differentiation of MC3T3-E1 cells.
Methods:
Differentially expressed miRNA was screened by miRNA Microarray. miR-7031-5P overexpression and knockdown MC3T3-E1 cell models were constructed by transfecting miR-7031-5P mimics and inhibitor. Alizarin red staining (ARS) assay was used to observe the formation of mineralized nodules in MC3T3-E1 cells. ALP activity was detected by using ALP detection kit. Western blot assay was used to examine the changes in osteogenic differentiation-related proteins. The relationship between miR-7031-5P and Wnt7a was revealed by dual luciferase report experiments.
Results:
We found that miR-7031-5P was up-regulated in MC3T3-E1 cells after Hyp treatment. The results indicated that compared with the untreated group, Hyp promoted the formation of mineralized nodules and the alkaline phosphatase (ALP) activity of MC3T3-E1 cells via overexpressing miR-7031-5P. Besides, elevated miR-7031-5P increased OPN, COL1A1, and Runx2 mRNA expression. More importantly, Wnt7a was identified as the downstream target gene of miR-7031-5P promoting osteogenic differentiation of MC3T3-E1 cells.
Conclusions:
Hyp up-regulated miR-7031-5P to promote osteogenic differentiation of MC3T3-E1 cells by targeting Wnt7a.
