慢病毒介导microRNA-214在成牙骨质细胞分化过程中的作用

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Abstract Objective: To construct a silencing plasmid of microRNA-214 and study the functional role of microRNA-214 in the differentiation and mineralization of cementoblasts. Methods: The cementoblasts was cultured in normal condition and expression level of the cementoblastic markers was test during the differentiation and mineralization induction period. Silencing lentiviral vector of microRNA-214 was constructed and then transfected into mouse cementoblasts (OCCM-30 cell lines). Fluorescence microscope was used to detect transfection ability and silencing effects of the construct. Alizarin Red and Von kossa staining was used to test the mineralization ability of cementoblasts. Real-time PCR was used to demonstrate the differentiation ability of cementoblasts. Results: Real-time PCR results showed that the relative expression of cementoblast differentiation marker ALP, Runx2, BSP, OCN, Col1a and CAP mRNA increased gradually as time goes on. The blank group, negative control group and microRNA-214 group infection ratio of silent carrier reached more than 90% cementum cells. Compared with blank group and negative control group, the expression of microRNA-214 in the microRNA-214 silencing group decreased 5 times. The results showed that the microRNA-214 silencing vector played a good role in silence. Compared with blank group and negative control group, microRNA-214 silencing group could significantly increase the proliferation rate of cementoblasts. Compared with blank group and negative control group, microRNA-214 silencing group, the number of mineralized nodules increased and the morphology becamed larger. Cemen-toblast differentiation markers ALP, Runx2, OCN, Col1a, BSP and CAP expression were significantly increased in microRNA-214 silencing group in silence. Conclusion: The research demonstrats that silencing lentiviral vector of MicroRNA-214 can promote the differentiation and mineralization ability of cementoblasts.

Key words： microRNA-214 lentivirus dental cementum cell differentiation tooth remineralization

Received: 24 July 2016

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	SUN Fucai
	LIU Xueting
	CAI Zhibin
	ZHANG Jinhua.

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2017/V47/I3/187

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