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目的研究HBV对肝癌细胞中AKR1C1基因表达的影响,并对其作用机制进行初步的探讨。方法RT-PCR和Real-time PCR检测HepG2.2.15和HepG2细胞中AKR1C1基因的差异表达情况;构建AKR1C1基因启动子虫荧光素酶报告质粒,并分别与HBV、HBx、HBs、HBp、HBc的表达质粒共转染HepG2细胞,双荧光素酶检测系统检测虫荧光素酶活性。结果AKR1C1基因在HepG2.2.15细胞中的表达高于HepG2细胞1.5倍;在HBV表达质粒与AKR1C1基因启动子虫荧光素酶报告质粒共转染的HepG2细胞中,虫荧光素酶的活性明显增强,为对照组3.37倍;相对HBs、HBp、HBc,HBx升高虫荧光素酶的活性的作用最强。结论HBV可以增强AKR1C1基因启动子的转录活性,促进AKR1C1在肝癌细胞中的表达,HBx在这一转录激活当中发挥重要作用。
Objective To study the effect of HBV on AKR1C1 gene expression in hepatocellular carcinoma cells and to investigate its mechanism. The differential expression of AKR1C1 gene in HepG2.2.15 and HepG2 cells was detected by RT-PCR and Real-time PCR. The luciferase reporter plasmid of AKR1C1 promoter was constructed and the expression of HBV, HBx, HBs, HBp and HBc Plasmids were co-transfected into HepG2 cells and luciferase activity was detected by dual luciferase assay. Results The expression of AKR1C1 gene in HepG2.2.15 cells was 1.5 times higher than that in HepG2 cells. The luciferase activity of HepG2 cells co-transfected with the HBV expression plasmid and the promoter of luciferase reporter gene AKR1C1 gene was significantly increased, 3.37 times as much as the control group; HBs, HBc, HBc and HBx increased the luciferase activity most strongly. Conclusion HBV can enhance the transcriptional activity of AKR1C1 promoter and promote the expression of AKR1C1 in hepatocellular carcinoma cells. HBx plays an important role in this transcriptional activation.