【摘要】 目的：研究脂肪与肥胖相关基因(FTO)对肝细胞的增殖效果。方法：构建FTO慢病毒过表达载体FTO-OE、慢病毒干扰载体FTO-sh,转染到肝细胞，western blot与qPCR分别分析其表达效果；CCK-8检测其对肝细胞抑制率；分析FTO对甘油及甘油三酯代谢的调控；qPCR检测肝细胞脂肪代谢相关基因的表达，脂肪分解相关基因(ATGL)、成脂分化基因(c/EBPβ)、脂肪酸从头合成相关基因(FAS)。结果：CCK-8结果显示，FTO-OE促进肝细胞增殖活性，FTO-sh抑制肝细胞增殖活性(P<0.05)。western blot与qPCR结果显示，FTO-OE能有效促进其表达，FTO-sh能有效沉默其表达，证实载体构建的有效性(P<0.05)。FTO-OE有效提高甘油及甘油三酯水平，FTO-sh降低甘油及甘油三酯水平(P<0.05)。qPCR结果显示，FTO-OE降低ATGL的表达、提高c/EBPβ与FAS的表达水平(P<0.05);FTO-sh提高ATGL的表达、降低c/EBPβ与FAS的表达水平(P<0.05)。结论：FTO具有促进肝细胞增殖的效果。更多还原

【Abstract】 Objective:To study the effect of FTO in regulating the proliferation of hepatocyte.Methods:Lentiviral vector FTO-OE and FTO-sh were constructed and were transduced in hepatocyte. The efficiency was examined by western blot and qPCR. The proliferation and viability in hepatocyte was used by CCK-8 assay. The glycerol and triglyceride level in hepatocyte was analyzed. The fat metaolism-related gene, including ATGL,c/EBPβ, FAS,were evaluated by qPCR assay.Results:CCK-8 result shown hepatocyte proliferation was regulated by FTO. Western blot and qPCR results demonstrated that FTO-OE vector increased the level of FTO and FTO-sh vector reduced the level of FTO(P<0.05).FTO-OE increased the level of glycerol and triglyceride and FTO-sh reduced them(P<0.05).qPCR results demonstrated that FTO-OE redued the level in ATGL, increased the level of c/EBPβ and FAS(P<0.05).FTO-sh results demonstrated the reverse the effect in genes level(P<0.05).Conclusion:FTO increased the proliferation in hepatocyte.更多还原