论文部分内容阅读
目的探讨HBx小发夹RNA(shRNA)对HepG2.2.15细胞中基质金属蛋白酶2(MMP-2)表达的影响。方法采用psiHBV/X质粒转染HepG2.2.15细胞,反转录PCR评估沉默效率;MTT法检测转染后HepG2.2.15细胞增殖情况;实时定量PCR(qRT-PCR)检测MMP-2 mRNA表达;Western blot法检测MMP-2蛋白表达的变化。结果反转录PCR检测HBx基因的沉默效率为53.6%;MTT检测结果显示转染24、48、72 h后HepG2.2.15细胞的增殖受抑制,与对照组比较,差异有统计学意义(P<0.05);psiHBV/X质粒转染HepG2.2.15细胞后,qRT-PCR和Western blot检测结果显示,HepG2.2.15细胞中MMP-2基因的mRNA水平和蛋白水平表达分别有不同程度的下调,与对照组比较差异有统计学意义(P<0.05)。结论 RNA干扰技术抑制HepG2.2.15细胞中HBx基因的表达,可抑制细胞增殖,下调HepG2.2.15细胞中MMP-2的表达。
Objective To investigate the effect of HBx small hairpin RNA (shRNA) on the expression of matrix metalloproteinase 2 (MMP-2) in HepG2.2.15 cells. Methods HepG2.2.15 cells were transfected with psiHBV / X plasmid and the silencing efficiency was evaluated by reverse transcription PCR. The proliferation of HepG2.2.15 cells was detected by MTT assay. The expression of MMP-2 mRNA was detected by real-time quantitative PCR (qRT-PCR) blot assay MMP-2 protein expression changes. Results The silencing efficiency of HBx gene by RT-PCR was 53.6%. The results of MTT assay showed that the proliferation of HepG2.2.15 cells was inhibited at 24, 48 and 72 hours after transfection. Compared with the control group, the difference was statistically significant (P < 0.05). The results of qRT-PCR and Western blot showed that the expression of MMP-2 mRNA and protein in HepG2.2.15 cells were down-regulated in different degrees compared with the control The difference was statistically significant (P <0.05). Conclusion RNAi can inhibit the expression of HBx gene in HepG2.2.15 cells, inhibit cell proliferation and down-regulate the expression of MMP-2 in HepG2.2.15 cells.