KAT2B对脂肪细胞分化调控的分子机制研究

项目名称： KAT2B对脂肪细胞分化调控的分子机制研究

项目编号： No.31201029

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 遗传学与生物信息学、细胞生物学

项目作者： 时文涛

作者单位： 天津医科大学

项目金额： 23万元

中文摘要： 脂肪细胞分化与肥胖的发生、发展关系密切。组蛋白乙酰转移酶KAT2B在基因转录调控中起重要作用，但该基因是否及如何参与脂肪细胞分化的调控尚存疑问。我们前期应用3T3-L1前体脂肪细胞研究发现：①KAT2B表达量在细胞诱导分化后升高；②KAT2B与PPARγ1间存在直接的蛋白交互作用；③KAT2B表达敲低稳定细胞株分化程度显著降低；④ KAT2B表达敲低引起CEBPα基因和几种脂肪因子表达的下调。上述结果提示，KAT2B参与了脂肪细胞分化的调控，其作用的分子机制值得我们进一步研究。本课题将从KAT2B这个表观遗传水平的调控分子出发，①用ChIP分析与其结合的DNA调控序列；②用PCR Array鉴定其参与调控的下游脂肪细胞分化相关基因；③根据前期基因型交互作用结果用Co-IP筛选与其结合的转录因子。课题的成果将完善脂肪细胞分化转录调控网络，为肥胖等疾病的预防和治疗提供理论依据和新靶点。

中文关键词： Kat2b；3T3-L1 细胞；Foxo1；转录调控；

英文摘要： Adipocyte differentiation contributes significantly to the development of obestiy. KAT2B, a key component of the histone acetyl transferase (HAT) complex, has profound effects on transcriptional regulations. However, it is unclear if KAT2B had a direct impact on adipocyte differentiation. Our previous studies showed that the expression of KAT2B increased significantly after 3T3-L1 preadipocytes differentiations were induced; Co-IP assays showed that KAT2B interacted directly with PPARγ1; Compare with untreated 3T3-L1 cells, KAT2B knockdown stable cell line had impaired differentiation rather than proliferation; We also found that the KAT2B knockdown downregulated CEBPα and several adipokines. These results suggest that KAT2B plays an importnant role in preadipocyte differentiations, obviously, the mechanism needs further investigations. Specific aims of our project is to investigate plausible roles of KAT2B as an HAT component and an epigenetic regulator of adipocyte differentiaions, we would like to 1) conduct ChIP assays in differentiated 3T3-L1 adipocytes to determine the location of DNA binding sites of KAT2B; 2) manipulate KAT2B expressions by Lentiviral ORF/shRNA systems, detect target gene expression profiles by Real-time PCR arrays; and 3) According to our previous genotype interaction results, find and

英文关键词： Kat2b；3T3-L1 cells；Foxo1；transcription regulation；