Background: During evolution,genomes gradually acquire novel genetic elements,including novel genes that are species/***'s known that some novel genes substantially contribute to the increasing functional and phen...
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Background: During evolution,genomes gradually acquire novel genetic elements,including novel genes that are species/***'s known that some novel genes substantially contribute to the increasing functional and phenotypic ***,there's no genome-wide investigation of their features and regular patterns contributing to organismal ***,we give a comprehensive analysis about the structural and functional features of novel genes and explore their roles in the formation of organismal complexity.
目的:建立由于油酸大量摄入造成的HepG2细胞脂肪变性模型;发现HepG2细胞发生脂肪变性前、后转录因子DNA结合活性的差异。方法:将HepG2细胞在含2mmol/L油酸的培养基中培养24h,通过油红O染色鉴定细胞中的甘油三酯含量,并确定发生脂肪变性的程度。分别提取对照组和实验组的核蛋白,利用转录因子富集技术cat TFRE(a concatenated tandem array of the consensus transcription factor response element),同时运用深度覆盖的蛋白质组学手段,对HepG2细胞脂肪变性前、后的转录因子变化进行动态描绘和定量分析。最后,利用IPA(integrated pathway analysis)对DNA结合活性发生改变的转录因子进行功能分析。结果:对照组总共鉴定到170种转录因子,实验组总共鉴定到190种转录因子,两组总共鉴定到208种转录因子。HepG2细胞发生脂肪变性后,DNA结合活性发生变化的转录因子有101种(重复实验间差异小于2倍,不同处理间差异大于2倍),DNA结合活性加强的有67种,减弱的有34种。其中,调节糖代谢、脂代谢的关键转录因子MLX、MLXIPL的DNA结合活性减弱;NF-κB1的DNA结合活性增加,暗示了NF-κB介导的炎症反应的激活。IPA分析的结果表明,油酸激活了HepG2细胞内NRF2介导的氧化应激反应,促进了基因表达(gene expression)、细胞增殖(cell proliferation)、细胞分化(differentiation of cell)、细胞周期(cell cycle)及细胞存活(cell survival)的进程。结论:油酸诱导HepG2细胞脂肪变性会引起细胞内糖、脂代谢的紊乱,诱发炎症反应,并引起氧化损伤,但是在转录水平上,细胞通过加强脂类代谢基因的表达、减少糖类向脂肪酸的转化、促进细胞增殖及激活NRF2介导的氧化应激反应等,最终促进了HepG2细胞的存活。
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