脂肪与肥胖相关基因FTO对肝细胞增殖效果研究

Study of the Hepatocyte Proliferation Regulated by FTO

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。

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/EBPβand 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.