Zhuo Dong, Ning Jin-Zhuo, Zhang Jing-Wei, Chen Yang, Cheng Li, Ba Zheng
Department of Urology, Yijishan Hospital, Wannan Medical College, Wuhu, Anhui 241000, China.
Department of Urology, Renmin Hospital of Wuhan University, Wuhan, Hubei 430061, China.
Zhonghua Nan Ke Xue. 2020 Jun;26(6):499-504.
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.
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-203a and PTEN by qRT-PCR and Western blot, respectively.
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-203a markedly decreased. Silencing lncRNA H19 enhanced the proliferation and inhibited the apoptosis of the GC-1 cells, and up-regulated the expression of microRNA-203a and down-regulated that of PTEN in the GC-1 cells.
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-203a /PTEN signaling pathway.
探讨长链非编码RNA(lncRNA)H19在体外培养的小鼠GC-1细胞中的表达及其对GC-1细胞增殖和凋亡的影响。
建立GC-1细胞体外缺氧复氧模型,采用qRT-PCR检测复氧损伤不同时间点GC-1细胞中lncRNA H19的表达。通过MTT法和流式细胞术检测沉默lncRNA H19对GC-1细胞增殖和凋亡的影响,采用Western blot检测GC-1细胞中凋亡相关蛋白Bax和caspase-3的表达,分别采用qRT-PCR和Western blot检测GC-1细胞中微小RNA-203a(miR-203a)和磷酸酶和张力蛋白同源物(PTEN)的表达。
随着复氧损伤时间的延长,GC-1细胞中lncRNA H19的表达显著增加,在缺氧3小时和复氧12小时达到峰值,而miR-203a的表达明显降低。沉默lncRNA H19可增强GC-1细胞的增殖并抑制其凋亡,上调GC-1细胞中miR-203a的表达并下调PTEN的表达。
lncRNA H19在体外培养的GC-1细胞中高表达,可能通过调节miR-203a/PTEN信号通路影响GC-1细胞的增殖和凋亡。