VPS13A在3T3-L1脂肪细胞分化过程中的表达及调控研究  

Expression and regulation of VPS13A in 3T3-L1 adipocyte differentiation

作　　者：刘瑾 黄昀 李超普 张桐 潘锦堃 季学涛 张许 李仲 LIU Jin;HUANG Yun;LI Chaopu;ZHANG Tong;PAN Jinkun;JI Xuetao;ZHANG Xu;LI Zhong(Key Laboratory of Rare Metabolic Disease,Department of Molecular Biology and Biochemistry,Nanjing Medical University,the Key Laboratory of Human Functional Genomics of Jiangsu Province,Nanjing 211166,China)

机构地区：[1]南京医科大学罕见代谢性疾病研究重点实验室,南京医科大学生物化学与分子生物学系,江苏省人类功能基因组重点实验室,江苏南京211166

出　　处：《南京医科大学学报（自然科学版）》2023年第8期1041-1046,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(32130050,3220110126);南京医科大学科技发展基金(2016NJMU004)。

摘　　要：目的:明确囊泡分选蛋白13A(vacuolar protein sorting 13 homolog A,VPS13A)在小鼠胚胎成纤维细胞(mouse embry-onic fibroblast cells,3T3-L1)分化过程中的表达水平;探索VPS13A对3T3-L1细胞的影响。方法:利用Western blot及Q-PCR法检测3T3-L1细胞在分化过程中VPS13A的表达,利用CRISPR/Cas9系统构建VPS13A稳定敲降细胞系,Western blot和油红染色检测VPS13A敲降后对3T3-L1细胞分化的影响。结果:VPS13A的表达在3T3-L1分化过程的早期降低,在分化的后期升高。敲降VPS13A后,3T3-L1细胞中脂滴增多,参与调控脂肪细胞分化基因的表达水平升高。结论:在3T3-L1细胞中VPS13A的表达水平在分化过程中被调控,敲降VPS13A可以促进3T3-L1细胞的分化成熟。Objective:To determine the expression level of vacuolar protein sorting 13 homolog A(VPS13A)during cell differentiation of 3T3-L1 and to explore the effects of VPS13A on 3T3-L1 cells.Methods:Western blot and Q-PCR were used to detect the expression changes of VPS13A during the differentiation of 3T3-L1 cells,and CRISPR(clustered regularly interspaced short palindromic repeats)/Cas9(CRISPR associated with 9)system to establish stable VPS13A knowdown cell lines.Finally,the effect of VPS13A knockdown on 3T3-L1 cell differentiation was detected by Western blot and Oil red staining.Results:The expression of VPS13A was downregulated in the early stage and upregulated in the later stage of 3T3-L1 cell differentiation.Lipid droplets and the expression levels of genes involved in differentiation were increased in VPS13A knockdown 3T3-L1 cells.Conclusion:VPS13A expression is modulated by cell differentiation status in 3T3-L1 cells.Knockdown of VPS13A enhances the differentiation of 3T3-L1 cells.

关 键 词：VPS13A 3T3-L1细胞 脂滴 脂肪细胞分化 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]