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目的利用siRNA技术探讨HBx基因沉默对肝癌细胞HepG2.2.15中MMP-9表达的影响。方法将针对HBx基因的干扰质粒载体pSIHBV/X转染肝癌细胞HepG2.2.15,用Real-time PCR法检测HBx与MMP-9 mRNA表达;Western blot方法检测MMP-9蛋白表达。结果 Real-time PCR检测结果显示,转染24 h、48 h、72 h后实验组HepG2.2.15细胞中HBx基因mRNA相对表达水平分别为121.13±8.72、83.17±7.93、56.33±6.17,与对照组相比(137.28±12.33)差异有统计学意义(P<0.01);24 h、48 h、72 h实验组HepG2.2.15细胞中MMP-9基因mRNA相对表达水平分别为83.27±6.17、69.37±8.26、58.18±4.77,与对照组相比(131.27±16.28)差异有统计学意义(P<0.01);Western blot检测结果表明,24 h、48 h实验组HepG2.2.15细胞中MMP-9蛋白相对表达水平分别为0.357±0.025、0.218±0.051,与对照组相比(0.475±0.073)差异有统计学意义(P<0.05)。结论利用HBx siRNA干扰质粒沉默HepG2.2.15细胞中HBx基因表达,可下调与肿瘤细胞迁移相关的MMP-9基因mRNA水平和蛋白水平的表达。
Objective To explore the effect of HBx gene silencing on the expression of MMP-9 in HepG2.2.15 cells by siRNA. Methods HBx gene was transfected into HepG2.2.15 cells with pSIHBV / X vector. The expression of HBx and MMP-9 mRNA was detected by Real-time PCR. The protein expression of MMP-9 was detected by Western blot. Results Real-time PCR results showed that the relative expression levels of HBx mRNA in HepG2.2.15 cells after transfection for 24 h, 48 h and 72 h were 121.13 ± 8.72, 83.17 ± 7.93 and 56.33 ± 6.17, respectively. Compared with the control group (137.28 ± 12.33) (P <0.01). The relative expression levels of MMP-9 mRNA in HepG2.2.15 cells at 24 h, 48 h and 72 h were 83.27 ± 6.17 and 69.37 ± 8.26 , 58.18 ± 4.77, respectively, compared with the control group (131.27 ± 16.28) (P <0.01). The results of Western blot showed that the relative expression of MMP-9 protein in HepG2.2.15 cells in 24 h and 48 h experimental groups The levels were 0.357 ± 0.025 and 0.218 ± 0.051, respectively, which were significantly different from the control group (0.475 ± 0.073) (P <0.05). Conclusion HBx siRNA silencing plasmid expression of HBx gene in HepG2.2.15 cells can down-regulate the expression of MMP-9 mRNA and protein in tumor cells.