Chinese Journal of Tissue Engineering Research ›› 2018, Vol. 22 ›› Issue (24): 3792-3797.doi: 10.3969/j.issn.2095-4344.0313

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Effect of OTX2 gene on the proliferation and differentiation of hFOB1.19 osteoblasts

Wang Jing1, 2, Tian Yu-lou2   

  1. 1the First Clinical Department of China Medical University, Shenyang 110122, Liaoning Province, China; 2Department of Orthodontics, School of Stomatology, China Medical University, Shenyang 110002, Liaoning Province, China
  • Received:2018-04-28
  • Contact: Tian Yu-lou, M.D., Professor, Department of Orthodontics, School of Stomatology, China Medical University, Shenyang 110002, Liaoning Province, China
  • About author:Wang Jing, Master, the First Clinical Department of China Medical University, Shenyang 110122, Liaoning Province, China; Department of Orthodontics, School of Stomatology, China Medical University, Shenyang 110002, Liaoning Province, China
  • Supported by:

    the Science and Technology Program of Shenyang, No. F15-199-1-55; the Science Research Project of Department of Education of Liaoning Province, No. LK201639

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Abstract:

BACKGROUND: Studying the expression and differentiation of osteoblasts contributes to understanding the mechanism of hereditary mandibular deformity at molecular level.
OBJECTIVE: To down-regulate the expression of OTX2 gene and investigate the effect of OTX2 gene on the proliferation and differentiation of hFOB1.19 osteoblasts by RNA interference.
METHODS: There were six groups: synthesized OTX2- siRNA1, OTX2-siRNA2, OTX2-siRNA3, GAPDH-siRNA, negative control, and blank control groups. Three target sequence siRNAs were designed and transfected into the osteoblasts. The OTX2 mRNA level in osteoblasts was detected by RT-PCR after transfection. The protein expression level was detected by western blot assay. The inhibition of OTX2-siRNA on osteoblast proliferation was evaluated by MTT assay. The alkaline phosphatase activity was detected by chemical colorimetry.
RESULTS AND CONCLUSION: OTX2 mRNA and protein expression was down-regulated after transfection. In the OTX2-siRNA group, the number of loating cells and cell proliferation inhibition rate were higher and the alkaline phosphatase activity was decreased compared with the control group. Our findings indicate that the chemically synthesized OTX2-siRNA can effectively restrain mRNA expression level of OTX2 gene in hFOB1.19 osteoblasts. Interference of OTX2 gene can inhibit the proliferation of hFOB1.19 osteoblasts, and down-regulated expression level of OTX2 gene can reduce the alkaline phosphatase activity of hFOB1.19 osteoblasts and inhibit their differentiation.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Osteoblasts, RNA Interference, Cell Proliferation, Tissue Engineering

CLC Number: