鸡CTCF基因真核表达载体的构建及其对脂类代谢作用的研究  

Construction of Chicken CTCF Gene Eukaryotic Expression Vector and Its Effect on Lipid Metabolism

作　　者：夏磊 魏志恒 裴军 刘世昊 周博 徐璐 郁建锋[1] 顾志良[1] XIA Lei;WEI Zhiheng;PEI Jun;LIU Shihao;ZHOU Bo;XU Lu;YU Jianfeng;GU Zhiliang(School of Biology and Food Engineering,Changshu Institute of Technology,Changshu 215500,China)

机构地区：[1]常熟理工学院生物与食品工程学院,江苏常熟215500

出　　处：《常熟理工学院学报》2025年第2期54-60,共7页Journal of Changshu Institute of Technology

基　　金：江苏省自然科学基金项目“RNA去甲基化酶FTO对鸡肝脏细胞脂类代谢和脂肪沉积作用的机制研究”(BK20191476)。

摘　　要：CCCTC结合因子(CCCTC-binding factor,CTCF)是一种多功能且高度保守的转录调节因子,参与转录调控、染色质结构维护、细胞分化等多种生物学过程.本研究通过RT-qPCR检测CTCF基因在鸡不同组织中的表达量,发现CTCF在脑和心脏中高表达.构建的CTCF真核过表达载体转染鸡肝癌细胞系LMH(Leghorn Male Hepatoma,LMH)后,成功表达CTCF-Myc融合蛋白.油红O染色和比色法检测显示,过表达CTCF后,LMH细胞脂滴沉积和总甘油三酯(TG)显著增加(P<0.01),磷脂酸磷酸酶1(LIPIN1)基因表达下调(P<0.01),固醇调节元件结合蛋白1(SREBP1)、脂肪酸合酶(FASN)和乙酰辅酶A羧化酶α(ACACA)基因表达显著上调(P<0.05或P<0.01).结果表明,CTCF基因可能通过调控脂质代谢相关基因表达促进脂质沉积.The CCCTC-binding factor(CTCF)is a multifunctional and highly conserved transcriptional regulator involved in processes such as transcriptional control,chromatin structure maintenance,and cellular differentiation.This study utilized RT-qPCR to analyze CTCF gene expression across various chicken tissues,revealing elevated expression levels in the brain and heart.A eukaryotic overexpression vector containing the CTCF gene was constructed and transfected into the Leghorn male hepatoma(LMH)cell line,successfully expressing the CTCF-Myc fusion protein.Oil Red O staining and colorimetric analysis demonstrated that CTCF overexpression in LMH cells significantly increased lipid droplet accumulation and total triglyceride(TG)levels(P<0.01).Concurrently,expression of Lipin 1(LIPIN1)was significantly downregulated(P<0.01),while expressions of Sterol Regulatory Element-Binding Protein 1(SREBP1),Fatty Acid Synthase(FASN),and Acetyl-CoA Carboxylase Alpha(ACACA)were significantly upregulated(P<0.05 or P<0.01).These results suggest that CTCF gene may promote lipid accumulation by regulating the expression of genes involved in lipid metabolism.

关 键 词：CCCTC结合因子 鸡肝癌细胞 脂滴沉积 甘油三酯 脂类代谢相关基因 

分 类 号：S831.2[农业科学—畜牧学]