摘要
目的探究乙型肝炎病毒X(HBX)蛋白过表达对肝癌细胞脂代谢及CCAAT增强子结合蛋白α/固醇调节元件结合蛋白-1(CEBPα/SREBP-1)通路的影响。方法用HBX过表达质粒转染人肝癌细胞,细胞计数试剂盒(CCK-8)检测细胞增殖,油红O染色检测肝癌细胞中脂滴堆积情况,蛋白质印迹法(Westernblot)检测脂代谢相关基因C/EBPa、SREBP-1和脂肪酸合成酶(FASN)的表达水平;CCK-8法、油红O染色和Westernblot检测C/EBPa在过表达对HBX调节人肝癌细胞脂代谢和增殖的影响。采用单因素方差和t检验分析。结果HBX表达增加显著促进人肝癌细胞脂质堆积(P<0.05),且增加脂质代谢的重要调节因子C/EBPα(0.15比0.38,t=1.351,P<0.05)和SREBP的表达水平(0.25比0.29,t=1.252,P<0.05)。C/EBPα过表达进一步加强了HBX对人肝癌细胞C/EBPα/(0.22比0.39,t=1.343,P<0.05)、SREBP-1(0.19比0.36,t=1.481,P<0.05)和FASN的表达(0.24比0.42,t=1.422,P<0.05)。结论HBX和C/EBPα相互作用,并通过影响下游基因SREBP-1的表达,从而影响肝癌细胞的增殖和脂质生成。
Objective To investigate the effects of hepatitis B virus X(HBX)protein overexpres-sion on lipid metabolism and CCAAT enhancer binding proteinα/sterol regulatory element binding protein-1(CEBP-α/SREBP-1)pathway in hepatocellular carcinoma cells.Methods Human liver cancer cells were transfected with HBX overexpression plasmid,cell proliferation was detected by cell counting kit-8(CCK-8)method,lipid accumulation in liver cancer cells was detected by oil red O staining,and the ex-pression levels of lipid metabolis-related genes C/EBPa,SREBP-1 and fatty acid synthetase(FASN)were detected by Western blotting.The effects of C/EBPa overexpression on the regulation of HBX on lipid me-tabolism and proliferation of human hepatoma cells were detected by CCK-8 assay,oil red O staining and Western blotting.Univariate variance and t test was used for statistical analysis.Results Increased HBX expression significantly promoted lipid accumulation in human hepatoma cells(P<0.05).It also increased the expression levels of C/EBp-α(0.15 vs.0.38,t=1.351,P<0.05)and SREBP(0.25 vs.0.29,t=1.252,P<0.05).Overexpression of C/EBp-αfurther enhanced the effect of HBX on C/EBp-α(0.22 vs.0.39,t=1.34,P<0.05),SEBP-1(0.19 vs.0.36,t=1.481,P<0.05)and FASN(0.24 vs.0.42,t=1.422,P<0.05).Conclusion HBX interacts with C/EBP-αand affects the proliferation and lipid produc-tion of hepatocellular carcinoma cells by influencing the expression of downstream gene SREBP-1.
作者
吴海俊
程国平
李辉华
邓海山
张永涛
Wu Haijun;Cheng Guoping;Li Huihua;Deng Haishan;Zhang Yongtao(Department of General SurgeryA,rmed Police Marine Police Corps Hospital,Jiaxing314000,China;Department of Pathology,Zhejiang Cancer Hospital,Hangzhou 310022,China)
出处
《中华实验外科杂志》
CAS
北大核心
2023年第7期1312-1314,共3页
Chinese Journal of Experimental Surgery
基金
2021年浙江省卫生健康委员会卫生健康科技计划基金(2021KY579)。
