CAJ | 학술논문

目的：研究GATA6基因沉默对肝癌Huh-7细胞凋亡的影响，同时探讨可能的作用机制。方法：构建靶向GATA6基因的慢病毒干扰载体，用流式细胞术确定转染效率，用RT-PCR和Western blotting法检测其对人类肝细胞癌细胞株Huh-7的干扰效果，流式细胞术检测细胞凋亡比例， Western blotting法检测各转染组Huh-7细胞NF-κB及Bcl-2蛋白表达。结果：靶向GATA6基因的慢病毒干扰载体感染肝癌Huh-7细胞后，转染效率为57.4％。GATA6在mRNA和蛋白水平的表达明显下降。 GATA6沉默的肝癌Huh-7细胞凋亡比例增加（ P＜0.05），NF-κB和Bcl-2蛋白水平表达降低。结论：利用慢病毒载体干扰技术沉默GATA6基因的表达可以明显促进Huh-7细胞凋亡，其机制可能通过NF-κB信号通路调节凋亡相关蛋白的表达，进而影响细胞的凋亡。靶向GATA6的RNA干扰技术在肝癌的基因治疗中具有一定的研究价值。
목적：연구GATA6기인침묵대간암Huh-7세포조망적영향，동시탐토가능적작용궤제。방법：구건파향GATA6기인적만병독간우재체，용류식세포술학정전염효솔，용RT-PCR화Western blotting법검측기대인류간세포암세포주Huh-7적간우효과，류식세포술검측세포조망비례， Western blotting법검측각전염조Huh-7세포NF-κB급Bcl-2단백표체。결과：파향GATA6기인적만병독간우재체감염간암Huh-7세포후，전염효솔위57.4％。GATA6재mRNA화단백수평적표체명현하강。 GATA6침묵적간암Huh-7세포조망비례증가（ P＜0.05），NF-κB화Bcl-2단백수평표체강저。결론：이용만병독재체간우기술침묵GATA6기인적표체가이명현촉진Huh-7세포조망，기궤제가능통과NF-κB신호통로조절조망상관단백적표체，진이영향세포적조망。파향GATA6적RNA간우기술재간암적기인치료중구유일정적연구개치。
AIM:To explore the effect of GATA6 gene silencing on apoptosis of hepatocellular carcinoma Huh-7 cells.METHODS:RNA interference vectors of the target gene GATA6 mediated by lentivirus were constructed in vitro to transfect the hepatocellular carcinoma cell line Huh-7.The apoptotic rate of transfected cells was measured by flow cytome-try.The protein expression of GATA6, NF-κB and Bcl-2 in transfected cells was determined by Western blotting.RE-SULTS:The transfection efficiency was 57.4%.The mRNA and protein expression of GATA6 reduced significantly after the carcinoma cell line Huh-7 being transfected by RNA interference vectors mediated by lentivirus.The apoptotic rate of the carcinoma cells with silent GATA6 gene was significantly increased (P<0.05).The protein expression levels of NF-κB and Bcl-2 were also significantly decreased.CONCLUSION: Lentiviral vector-mediated RNA interference of GATA6 has an inhibitory effect on the expression of the gene itself, and promotes the apoptosis of hepatocellular carcinoma cells.Regu-lation of the apoptosis-related protein expression by the NF-κB signaling to influence the proliferation and apoptosis of hepa-tocellular carcinoma cells might be one of the possible mechanisms.