Abstract:

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.

Key words: zinc-finger protein 260, mouse embryo osteoblast precursor cells, osteogenic differentiation

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。

关键词: 锌指蛋白260, 小鼠胚胎成骨细胞前体细胞, 成骨分化

CLC Number: 

• R782