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研究微重力对COL1A1(Ⅰ型胶原α1链基因)启动子活性的影响,探讨微重力对成骨细胞相关基因表达影响的作用机制.将长为3.6 kb COL1A1启动子双酶切,获得不同长度的启动子片段,并与报告基因EGFP(增强型绿色荧光蛋白)连接,转染ROS17/2.8细胞,用G418筛选,得到稳定转染COL1A1-EGFP基因的ROS17/2.8细胞株.利用回转器模拟微重力效应,体外培养条件下,观察各细胞株报告基因的表达情况.结果显示细胞在模拟微重力下培养24,48 h后,报告基因EGFP和Ⅰ型胶原的表达升高,表明COL1A1启动子活性增强.说明短期模拟微重力条件下,成骨细胞能通过增强COL1A1启动子活性,代偿性提高Ⅰ型胶原的表达.
To investigate the effect of microgravity on the promoter activity of COL1A1 (type Ⅰ collagen α1 chain gene) and to explore the mechanism of microgravity on osteoblast-related gene expression, double-digesting the 3.6 kb COL1A1 promoter to obtain different length Promoter fragment and linked with reporter gene EGFP (enhanced green fluorescent protein), transfected into ROS17 / 2.8 cells and screened by G418 to get ROS17 / 2.8 cell line stably transfected with COL1A1-EGFP gene.Micro-gravity Effect and in vitro culture conditions, the expression of reporter gene was observed.The results showed that the expression of EGFP and typeⅠcollagen increased after cells under simulated microgravity for 24,48 h, indicating that COL1A1 promoter activity was enhanced .Conclusion Under the conditions of short-term simulated microgravity, osteoblasts can compensatory increase the expression of type I collagen by enhancing the activity of COL1A1 promoter.