CAJ | 학술논문

目的：研究 HBx干扰质粒pSIHBV/X对肝癌细胞HepG2．2．15中VEGF表达的影响。方法将针对HBx基因的干扰质粒载体pSIHBV/X转染HepG2．2．15细胞，用Real-time PCR检测转染前后细胞 HBx、VEGF基因 mRNA 表达，Western blot法检测细胞 VEGF蛋白表达。结果Real-time PCR检测结果显示，转染后24 h、48 h、72 h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±7．930）、（56．33±6．17）；24 h、48 h、72 h实验组HepG2．2．15细胞中 VEGF基因 mRNA 相对表达水平分别为（76．26±7．16）、（59．17±6．420）、（42．17±4．33）；Western blot检测结果显示，24 h、48 h实验组 HepG2．2．15细胞中VEGF蛋白相对表达水平分别为（0．357±0．025）、（0．218±0．051）。Western blot及RT-PCR结果显示，与对照组相比实验组中VEGF mRNA 及蛋白的表达量明显下调，差异有统计学意义（P＜0．05）。结论靶向HBx的siRNA干扰质粒能抑制 HepG2．2．15细胞中VEGF mRNA和蛋白的表达。
목적：연구 HBx간우질립pSIHBV/X대간암세포HepG2．2．15중VEGF표체적영향。방법장침대HBx기인적간우질립재체pSIHBV/X전염HepG2．2．15세포，용Real-time PCR검측전염전후세포 HBx、VEGF기인 mRNA 표체，Western blot법검측세포 VEGF단백표체。결과Real-time PCR검측결과현시，전염후24 h、48 h、72 h실험조 HepG2．2．15세포중 HBx기인mRNA상대표체수평분별위（121．13±8．72）、（83．17±7．930）、（56．33±6．17）；24 h、48 h、72 h실험조HepG2．2．15세포중 VEGF기인 mRNA 상대표체수평분별위（76．26±7．16）、（59．17±6．420）、（42．17±4．33）；Western blot검측결과현시，24 h、48 h실험조 HepG2．2．15세포중VEGF단백상대표체수평분별위（0．357±0．025）、（0．218±0．051）。Western blot급RT-PCR결과현시，여대조조상비실험조중VEGF mRNA 급단백적표체량명현하조，차이유통계학의의（P＜0．05）。결론파향HBx적siRNA간우질립능억제 HepG2．2．15세포중VEGF mRNA화단백적표체。
Objective To investigate the effect of HBx siRNA on VEGF gene expression of HepG2 .2 .15 cells .Methods Specific siRNA against HBx was transfected into HepG2 .2 .15 cell .Ex-pressions of HBx and VEGF mRNA before and after transfection were detected using Real-time PCR . Expression of VEGF protein was analyzed by western blot (WB) .Results Real-time PCR testing re-sults showed that the expressions of HBx mRNA were (121 .13 ± 8 .72) ,(83 .17 ± 7 .93) ,(56 .33 ± 6 .17) in experimental group cells after transfection of 24 h ,48h ,72 h .The expressions of VEGF mRNA were(76 .26 ± 7 .16) ,(59 .17 ± 6 .42) ,( 42 .17 ± 4 .33) in experimental group cells after trans-fection of 24 h ,48 h ,72 h .WB testing results showed that the expressions of VEGF protein were (0 .357 ± 0 .025) ,(0 .218 ± 0 .051) in experimental group cells after transfection of 24 h ,48 h .Results of WB and Real-time PCR demonstrated that compared with control group cells ,expression levels of VEGF mRNA and protein were obviously down-regulated after transfection with pSIHBV/X ,and there were statistical significantly differences (P<0 .05) .Conclusion The research demonstrates that inhibition the expression of HBx gene by RNA interference could down-regulate VEGF gene at mRNA and protein level in HepG2 .2 .15 cells .