<em>Hey1</em> 基因参与调控BMP9诱导的C3H10T1/2细胞成骨分化

doi:10.13523/j.cb.20150603

中国生物工程杂志

2015, Vol. 35

Issue (6): 14-20 DOI: 10.13523/j.cb.20150603

研究报告

Hey1 基因参与调控BMP9诱导的C3H10T1/2细胞成骨分化

唐大刚, 王淼, 张艳亮, 王孝林, 李贵强, 罗小辑

重庆医科大学附属第一医院重庆 400016

Hey1 Gene Is Involved in Regulating BMP9-induced Osteogenic Differentiation of C3H10T1/2 Cells

TANG Da-gang, WANG Miao, ZHANG Yan-liang, WANG Xiao-lin, LI Gui-qiang, LUO Xiao-ji

Orthopedic Department, the First Affiliate Hospital of Chongqing Medical University, Chongqing 400016, China

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摘要：

目的:观察发状分裂相关增强子 Hey1 在骨形态发生蛋白9(BMP9)诱导的小鼠间充质干细胞(MSCs)C3H10T1/2成骨分化中的作用。方法:包装 Hey1、BMP9以及GFP的过表达慢病毒,并分别作用于C3H10T1/2细胞,RT-PCR和Western blot检测 Hey1、BMP9以及GFP的慢病毒是否包装成功,碱性磷酸酶(ALP)检测早期成骨指标ALP的变化,茜素红S染色检测晚期成骨指标钙盐沉积,MTT检测 Hey1 对BMP9调控的C3H10T1/2细胞增值的影响,流式细胞术检测 Hey1 对BMP9调控的C3H10T1/2细胞周期的影响。结果: Hey1、BMP9以及GFP的慢病毒包装成功;在成骨分化早期,过表达 Hey1 基因可促进BMP9调控的C3H10T1/2细胞的成骨分化与增殖;在成骨分化晚期,过表达 Hey1 基因可促进BMP9诱导的C3H10T1/2细胞的成骨分化, 并将BMP9调控的C3H10T1/2细胞周期阻滞在G1期。结论: Hey1 基因可参与调控BMP9诱导的小鼠C3H10T1/2 细胞的早晚期成骨分化,并对细胞的增殖与周期有一定的影响。

关键词： 发状分裂相关增强子Hey1; 骨形态发生蛋白9; C3H10T1/2细胞; 成骨分化

Abstract:

Objective: To analysis the effect of Hey1 gene on BMP9-induced osteogenic differentiation of mesenchymal stem cells C3H10T1/2. Methods: Assemble lentivirus LV5-GFP, LV5-BMP9 and LV5-Hey1, the expression of GFP, BMP9 and Hey1 were detected by RT-PCR and Western blot in C3H10T1/2 cells. Then C3H10T1/2 cells was infected with LV5-GFP, LV5-Hey1 and/or LV5-BMP9 respectively, the early osteogenic marker ALP activity was detected by quantitative assay, the later osteogenic marker calcium deposition was detected by Alizarin Red S staining, the cell proliferation and cycles regulated by BMP9 were detected by MTT and flow cytometry assay. Results: LV5-GFP, LV5-BMP9 and LV5-Hey1 promoted the expression of GFP, BMP9 and Hey1. LV5-Hey1 can increase the early and later osteogenic differentiation of C3H10T1/2 cells induced by BMP9, and promote the cell proliferation in the early stage, but inhibit cell cycles in the later stage regulated by BMP9. Conclusion: Hey1 gene can influence BMP9-induced osteogenic differentiation and BMP9-regulated cell proliferation and cell cycles of C3H10T1/2 cells.

Key words: Hey1 gene BMP9 C3H10T1/2 cells Osteogenic differentiation

收稿日期: 2015-02-26 出版日期: 2015-06-25

ZTFLH:

Q254

基金资助:

国家自然科学基金资助项目(81272171)

通讯作者: 罗小辑 E-mail: cy2982@163.com

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引用本文:

唐大刚, 王淼, 张艳亮, 王孝林, 李贵强, 罗小辑. Hey1 基因参与调控BMP9诱导的C3H10T1/2细胞成骨分化[J]. 中国生物工程杂志, 2015, 35(6): 14-20.

TANG Da-gang, WANG Miao, ZHANG Yan-liang, WANG Xiao-lin, LI Gui-qiang, LUO Xiao-ji. Hey1 Gene Is Involved in Regulating BMP9-induced Osteogenic Differentiation of C3H10T1/2 Cells. China Biotechnology, 2015, 35(6): 14-20.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20150603 或 https://manu60.magtech.com.cn/biotech/CN/Y2015/V35/I6/14

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