核酶基因抑制肝癌细胞低氧诱导因子-1α表达的实验研究

Inhibitory effects of hammerhead ribozyme on expression of hypoxia inducible factor 1α in hepatocarcinoma cells

目的探讨靶向HIF-1α核酶基因对肝癌细胞HIF-1α表达的调控。方法采用脂质体介导的方法，将靶向HIF-1α核酶基因真核表达载体转染肝癌细胞Hep3B2，并予低氧条件诱导。于转染后48h采用Western Blotting检测Hep3B2细胞中HIF-1α蛋白的表达水平；荧光报告基因方法检测HIF-1转录活性。结果Hep3B2细胞低氧诱导后，HIF-1α蛋白表达水平、HIF-1转录活性增高（1．0±0．02），转染核酶基因400μmol／L后48h，Hep3B2细胞低氧诱导的HIF-1α蛋白表达水平明显下降，HIF-1转录活性下调（0．12±0．025，P〈0．05）。结论核酶基因可特异性抑制低氧诱导的肝癌细胞HIF-1α表达，降低其转录活性。

Objective To explore the inhibition of hypoxia inducible factor-1α （HIF-1α） in hepatocarcinoma cells by hammerhead ribozyme. Methods anti-HIF-1α hammerhead ribozyme （RZ） eukaryotic expression vector was introduced into hepatocarcinoma cells Hep3B2 by lipofectin mediation. Western blotting and luciferase assay were used to detect the expression levels of HIF-1α protein and HIF-1 transcription activity in Hep3B2. Results The expression of HIF-1α in Hep3B2 cells and its transcription activity under hypoxia was higher than those under normoxia. After RZ transfection, the expression of HIF-1α protein and its transcription activity were down-regulated in the Hep3B2 cells in a dose-dependent manner. Conclusion anti-HIF-1α hammerhead ribozyme gene can inhibit the expression of HIF-1α and reduce its transcription activity in hepatocarcinoma cells.