Zfp260对MC3T3-E1细胞体外成骨分化及增殖、迁移影响的实验研究  

Experimental study on the effect of Zfp260 on osteogenic differentiation,proliferation and migration of MC3T3-E1 cells in vitro

作　　者：吴军 王佐林[1] WU Jun;WANG Zuolin(Shanghai Engineering Research Center of Tooth Restoration and Regeneration&Tongji Research Institute of Stomatology&Department of Oral Implantology,Shanghai Tongji Stomatological Hospital and Dental School,Tongji University,Shanghai 200072,China)

机构地区：[1]上海市同济口腔医院口腔种植科,同济大学口腔医学院,上海牙组织修复与再生工程技术研究中心,同济大学口腔医学研究所,上海200072

出　　处：《口腔颌面外科杂志》2024年第6期421-426,共6页Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(81600836)。

摘　　要：目的:探究锌指蛋白260(zinc-finger protein 260,Zfp260)对小鼠胚胎成骨细胞前体细胞(mouse embryo osteoblast precursor cells,MC3T3-E1 cells)体外成骨分化及增殖、迁移的影响。方法:体外培养并诱导MC3T3-E1细胞成骨分化,实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测诱导7、14 d后Zfp260的表达量变化。用小干扰RNA(small interfering RNA,siRNA)转染MC3T3-E1细胞,并通过RT-qPCR测定Zfp260的敲降效率及敲降组与对照组碱性磷酸酶(alkaline phosphatase,ALP)、人骨形态发生蛋白2(bone morphogenetic protein 2,BMP2)等成骨生物标志物表达量的变化。通过Transwell、细胞划痕实验检测敲降Zfp260后MC3T3-E1细胞迁移能力的变化。通过CCK8实验检测敲降Zfp260后MC3T3-E1细胞增殖能力的变化。结果:体外诱导MC3T3-E1细胞成骨分化后,Zfp260的表达量明显上调(P<0.05)。使用siRNA敲降Zfp260后,ALP及BMP2的表达量明显下调(P<0.05)。Transwell及细胞划痕实验证明敲降Zfp260后,MC3T3-E1细胞的迁移能力受到抑制。CCK8实验证明敲降Zfp260后,MC3T3-E1细胞的增殖能力增加。结论:Zfp260可促进MC3T3-E1细胞的成骨分化。Objective:To explore the effect of zinc‐finger protein 260(Zfp260)on the osteogenic differentiation,proliferation and migration of mouse embryo osteoblast precursor cells(MC3T3-E1 cells)in vitro.Methods:MC3T3-E1 cells were cultured in vitro and induced to differentiate into osteoblasts.The expression of Zfp260 was detected by real-time quantitative polymerase chain reaction(RT-qPCR)when it was induced for 7 days and 14 days.MC3T3-E1 cells were transfected with siRNA,and the knockdown efficiency of Zfp260 and the expression of alkaline phosphatase(ALP),human bone morphogenetic protein 2(BMP2)and other osteogenic biomarkers in knockdown group and control group were measured by RT-qPCR.The change of migration ability of MC3T3-E1 cells after knockdown of Zfp260 was detected by Transwell,cell scratch assay.The change of proliferation ability of MC3T3-E1 cells after knockdown of Zfp260 was detected by CCK8 experiment.Results:The expression of Zfp260 was significantly up-regulated after inducing osteogenic differentiation of MC3T3-E1 cells in vitro(P<0.05).After using siRNA to knock down Zfp260,the expression of ALP and BMP2 were significantly decreased(P<0.05).Transwell and cell scratch assays showed that the migration ability of MC3T3-E1 cells was inhibited after knocking down Zfp260.CCK8 experiment showed that the proliferation ability of MC3T3-E1 cells increased after knocking down Zfp260.Conclusion:Zfp260 can promote the osteogenic differentiation of MC3T3-E1 cells.

关 键 词：锌指蛋白260 小鼠胚胎成骨细胞前体细胞 成骨分化 

分 类 号：R782[医药卫生—口腔医学]