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目的探讨肝细胞癌高表达新基因BC047440在肝癌形成中的作用。方法通过 DOTAP~(TM)脂质体介导的转染技术,将BC047440基因反义真核表达载体导入HepG2肝癌细胞,经 C418筛选,获得稳定表达BC047440反义基因的HepG2肝癌细胞。采用半定量RT-PCR检测反义基因对转染细胞的内源性BC047440基因抑制效果,噻唑蓝(MTT)比色法观察转染细胞生长曲线及流式细胞术比较细胞周期的变化。结果转染BC047440反义基因的肝癌细胞内源性BC047440 基因受到有效抑制,细胞生长速度明显减慢,细胞周期中G_1,期细胞比例明显增高。结论肝细胞癌高表达基因BC047440可能是一个新的肝癌相关基因,在肝癌中的异常高表达可能对肝癌的发生发展具有一定的促进作用。
Objective To investigate the role of the novel high expression gene BC047440 in the development of hepatocellular carcinoma. Methods The antisense eukaryotic expression vector of BC047440 gene was transfected into HepG2 hepatoma cells by DOTAPTM liposome-mediated transfection. HepG2 hepatoma cells stably expressing BC047440 antisense gene were obtained through C418 selection. Semi-quantitative RT-PCR was used to detect the endogenous BC047440 gene-inhibitory effect of the antisense gene on the transfected cells. MTT colorimetric method was used to observe the growth curve of the transfected cells and flow cytometry to compare the cell cycle changes. Results The endogenous BC047440 gene in hepatoma cells transfected with BC047440 antisense gene was effectively inhibited, the growth rate of the cells was significantly slowed down, and the proportion of G 1 and phase cells in the cell cycle was significantly increased. Conclusion The high expression gene BC047440 of hepatocellular carcinoma may be a new liver cancer related gene. The abnormally high expression of BC047440 in hepatocellular carcinoma may play an important role in the development of hepatocellular carcinoma.