LncRNA H19/miR-203a/PTEN轴在小鼠GC-1细胞缺血再灌注损伤中的作用及影响机制

Role of the lncRNA H19/ microRNA-203a/PTEN axis in ischemia-reperfusion injury of mouse GC-1 cells and its mechanism

目的:探讨长链非编码RNA(lncRNA)H19在小鼠体外GC-1细胞的表达,以及其通过调控微小RNA(miRNA)-203a/磷酸酯酶与张力蛋白同源物(PTEN)轴对GC-1细胞凋亡和增殖的影响。方法:建立GC-1细胞体外缺氧复氧模型,qRT-PCR检测GC-1细胞不同复氧损伤时间点lncRNA H19的表达变化;MTT法、流式细胞术测定沉默lncRNA H19对GC-1细胞增殖和凋亡的影响;Western印迹检测沉默lncRNA H19对GC-1细胞凋亡相关蛋白Bax和caspase-3表达的影响;qRT-PCR和Western印迹分别测定沉默lncRNA H19后GC-1细胞miR-203a和PTEN的表达变化。结果:随着复氧损伤时间的增加,GC-1细胞lncRNA H19表达明显增加,在缺氧3 h/复氧12 h达到峰值,与此同时miR-203a表达明显降低。此外,沉默lncRNA H19增强了GC-1细胞的增殖能力并降低了其凋亡水平,增加了miR-203a表达水平并降低了PTEN表达水平,结果显著。结论:LncRNA H19在体外GC-1细胞中高表达,其可能通过调控miR-203a/PTEN信号途径改变GC-1细胞增殖及凋亡的能力。

Objective:To investigate the expression of long non-coding RNA(lncRNA)H19 in mouse GC-1 cells in vitro and its effects on the proliferation and apoptosis of GC-1 cells.Methods:We established an in vitro hypoxia-reoxygenation model in GC-1 cells and detected the expression of lncRNA H19 in the GC-1 cells at different time points of reoxygenation injury by qRT-PCR.We determined the effects of silencing lncRNA H19 on the proliferation and apoptosis of the GC-1 cells by MTT and flow cytometry,the expressions of apoptosis-related proteins Bax and caspase-3 in the GC-1 cells by Western blot,and the expressions of microRNA-203 a and PTEN by qRT-PCR and Western blot,respectively.Results:With the prolonging of the time of reoxygenation injury,the expression of lncRNA H19 was increased significantly in the GC-1 cells and peaked at 3-hour hypoxia and 12-hour reoxygenation,but that of microRNA-203 a markedly decreased.Silencing lncRNA H19 enhanced the proliferation and inhibited the apoptosis of the GC-1 cells,and up-regulated the expression of microRNA-203 a and down-regulated that of PTEN in the GC-1 cells.Conclusion:LncRNA H19 is highly expressed in GC-1 cells in vitro,which may influence the proliferation and apoptosis of GC-1 cells by regulating the microRNA-203 a/PTEN signaling pathway.