Impact of MicroRNA-103-3p on osteogenic differentiation of MC3T3-E1 at an early stage

doi:10.19438/j.cjoms.2018.04.003

Abstract

Abstract: PURPOSE:To investigate the effect of miR-103-3p on osteogenic differentiation of MC3T3-E1 cells at an early stage. METHODS: Osteogenic differentiation of MC3T3-E1 was induced in osteoblast induction medium. Runx2, Osx, ALP, miR-103-3p mRNA expression was detected by real-time PCR at day 0, 3, 5 and 7 following induction; Western blotting was conducted to show Runx2, Osx protein expression. Osteoblastic phenotype was estimated by alkaline phosphatase (ALP) staining. miR-103-3p mimics and mimics negative control were separately transiently transfected to MC3T3-E1 cells with lipofectamine 2000. The expression of miR-103-3p was determined by real-time PCR. Cell proliferation was detected by CCK-8 kit. The transfected MC3T3-E1 cells were induced into osteogenic differentiation. Real-time PCR was conducted to detect the levels of osteogenesis-related genes, such as Runx2, Osx and ALP. Western blotting was performed to confirm their expression on protein level at day 0, 3 and 7 following induction. ALP activity was shown by ALP staining. The data were analyzed by SPSS 19.0 software package. RESULTS: mRNA expression levels of Runx2, Osx and ALP were increasingly up-regulated in MC3T3-E1 with osteogenic induction. Protein expression levels of Runx2 and Osx were consistent with mRNA expression trends. miR-103-3p was suppressed during osteogenic differentiation of MC3T3-E1. After transient transfection of miR-103-3p mimics, the expression of miR-103-3p in the experimental group was significantly up-regulated (P<0.05). Cell proliferation was decreased. mRNA expression of Runx2 and ALP was markedly down-regulated (P<0.05). Runx2 protein expression was suppressed. After osteogenic induction for 3 and 7 days, the expression of Runx2, Osx and ALP on mRNA level was markedly down-regulated in the experimental group and so was the expression on protein level. CONCLUSIONS: miR-103-3p might suppress osteogenic differentiation of MC3T3-E1 cell at an early stage.

Key words: microRNA-103-3p, MC3T3-E1, Osteogenic differentiation

CLC Number:

Q254

LU Chen-pei, LI Hong-liang, OUYANG Ning-juan, LIN Yu-heng, SI Jia-wen, SHEN Guo-fang. Impact of MicroRNA-103-3p on osteogenic differentiation of MC3T3-E1 at an early stage[J]. China Journal of Oral and Maxillofacial Surgery, 2018, 16(4): 302-309.

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