microRNA-449通过调控SIRT1相关的脂质代谢途径抑制HepG2.2.15细胞增殖  

MicroRNA-449 suppresses proliferation of HepG 2.2.15 cell through regulation of SIRT1 related lipid metabolic pathway

作　　者：张文超[1] 王汉宁[1] 萧金丰[1] 陈开运[1] 贺轲[1] 段小鹏[1] 黄睿[1] 向国安[1] 

机构地区：[1]广东医学院教学医院广东省第二人民医院,广州510000

出　　处：《国际病毒学杂志》2015年第4期221-228,共8页International Journal of Virology

基　　金：基金项目：广东省自然科学基金（$2012010008274）;广东省科技计划项目（2011808701096）

摘　　要：目的 研究miRNA-449通过调控SIRT1相关脂质代谢途径抑制HepG 2.2.15增殖的作用机制.方法 使用Entrez Nucleotide database预测SIRT1是否为miRNA-449的靶基因.使用RT-PCR和western blot明确miRNA-449是否调控胆固醇调节元件结合蛋白1-脂肪生成-胆固醇生成代谢途径.BrdU实验明确miRNA-449对HepG 2.2.15有丝分裂的影响.miRNA-449模拟物或抑制剂转染HepG 2.2.15,观察miRNA-449对细胞增殖的影响.结果 miRNA-449通过下调SIRT1表达进一步抑制SREBP-1c及其下游的靶基因脂肪酸合酶（fatty acid synthase,FASN）和3-羟基-3-甲基戊二酰辅酶A还原酶（3-hydroxy-3-methylglutaryl CoA reductase,HMGCR）的表达来调控HepG 2.2.15细胞中脂肪生成及胆固醇合成.miRNA-449通过调节代谢途径抑制HepG 2.2.15细胞有丝分裂、增殖以及DNA合成.结论 miRNA-449可能通过下调SIRT1表达进一步抑制肝脏肿瘤的发生和发展.miRNA-449可能成为HBV感染相关肝细胞肝癌治疗的新靶点.Objective To explore the mechanism of MiRNA-449 suppressing proliferation of HepG 2.2.15 through regulation of SIRT1 related lipid metabolic pathway.Methods Entrez Nucleotide database was used to identify SIRT1 as the target genes of miRNA-449.Quantitative RT-PCR and western blot analysis were applied to confirme regulation of miRNAs on the SIRT1 and its downstream SREBP-lipogenesis-cholesterogenesis metabolic pathway in HepG 2.2.15 cells.The BrdU experiments,was used to confirm the effects of miRNA-449 on the mitosis of HepG 2.2.15 cells.HepG 2.2.15 cells were transfected with miRNA-449 mimics or inhibitors,and the effects of miRNA-449 on cell proliferation were assessed.Results The miRNA-449 could control lipogenesis and cholesterogenesis in HepG 2.2.15 cells by inhibiting SIRT1 and SREBP-1c expression and their down-regulating targeted genes,including fatty acid synthase （FASN） and 3-hydroxy-3-methylglutaryl CoA reductase （HMGCR）.The miRNA-449 repressed DNA synthesis,mitotic entry and proliferation of HepG 2.2.15 cell.Conclusions Restoration of miRNA-449 leads to suppression of SIRT1 expression and liver tumorigenesis.The newly identified miRNAs,miRNA-449 represents a novel targeting mechanism for HCC therapy.

关 键 词：miRNA-499 脂质代谢 肝癌 MiRNA-499 

分 类 号：R735.7[医药卫生—肿瘤]