肝细胞癌高表达新基因BC047440功能的研究

Primary study of the function of a novel gene BC047440 highly expressed in human hepatocellular carcinoma

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 initially study the function of a novel gene BC047440 highly expressed in human hepatoeeUular carcinoma. Methods We transfected HepG2 cell with the eukaryotic expression plasmid vector expressing BC047440 cDNA in the antisene orientation using DOTAP^TM liposome. The result of PCR exhibited that the recombinant plasmid containing BC047440 have integrated into the HepG2 cell. The effect of BC047440 antisene gene transfecrion on endogenous BC047440 mRNA levels was examined with RT-PCR, and the effects on the cell growth rate and the cell recycle were also examined by MTT colorimetry and Flow Cytometry, respectively. Results The hepatoma cells transfected with BC047440 anfisene gene exhibited a significant reduction in endogenous BC047440 mRNA levels, a lower cell growth speed and a higher ratio at G1 stage. Conclusion BC047440 is a novel HCC associated gene and highly involved in HCC development.