Hoxc6在胚胎间充质干细胞成骨分化中的作用

摘要

目的：研究homeobox c6(Hoxc6)在胚胎间充质干细胞成骨分化中的作用。方法 BMP4诱导小鼠胚胎间充质干细胞C3H10T1/2细胞体外成骨分化。逆转录病毒转染C3H10T1/2细胞,构建过表达Hoxc6的稳定转染细胞,进行Hoxc6获得性功能研究。慢病毒转染 C3H10T1/2细胞,构建基因敲除 Hoxc6的稳定转染细胞,进行Hoxc6丧失性功能研究。碱性磷酸酶活性实验检测成骨早期分化指标-碱性磷酸酶活性。实时荧光定量反转录PCR( real time reverse transcription PCR,qRT-PCR)检测成骨分化相关基因-碱性磷酸酶和骨钙素的表达。结果过表达Hoxc6明显增强C3H10T1/2细胞的碱性磷酸酶活性,在mRNA水平促进碱性磷酸酶和骨钙素的基因表达。基因敲除Hoxc6明显抑制C3H10T1/2细胞的碱性磷酸酶活性,明显抑制碱性磷酸酶和骨钙素的基因表达。结论 Hoxc6具有促进胚胎间充质干细胞体外成骨分化的功能。%Objective To investigate the role of homeobox c6 ( Hoxc6 ) in the osteogenic differentiation of embryonic mesenchymal cells. Methods BMP4 was used to induce the osteogenic differentiation of mouse embryonic mesenchymal cell line-C3H10T1/2. Retroviral HA-Hoxc6 was used to establish the stable cell and over-express Hoxc6 for Gain-of-function study. Lentiviral Hoxc6 shRNA was applied to establish the stable cell and knock-down of Hoxc6 for Loss-of-function study. The early marker of osteogenic differentiation-ALP activity was detected by alkaline phosphatase ( ALP) activity assay. The osteogenesis related genes expressions, Alp and osteocalcin ( Ocn) were examined by real time RT-PCR. Results Over-expression of Hoxc6 promoted the Alp activity and the expressions of Alp and Ocn in C3H10T1/2 cells. Knock-down of Hoxc6 inhibited the Alp activity and the expressions of Alp and Ocn in C3H10T1/2 cells. Conclusion Hoxc6 enhanced the osteogenic differentiation of embryonic mesenchymal cells in vitro.