表达谱芯片研究KCTD15基因在3TL1前体脂肪-第三军医大学学报.DOC

201312062 论著 表达谱芯片分析KCTD15对3T3-L1前体脂肪细胞的影响 张 鹏1，金康宣2，陈宇晟2，徐梓辉1 , 3（400037 重庆，第三军医大学新桥医院内分泌科1；中西医结合科3；100029 北京，中国科学院北京基因组研究所重大疾病基因组和个体化医疗实验室2） [摘要]目的 通过筛选分析3T3-L1前体脂肪细胞中KCTD15基因敲低后的差异表达基因研究其细胞生物学功能，探讨KCTD15基因与肥胖发生的关联。方法 ①运用RNA干扰技术在3T3-L1前体脂肪细胞中特异性敲低KCTD15基因表达，并用半定量PCR技术检测KCTD15基因敲低效率。②用Illumina表达谱芯片检测基因表达。③筛选差异表达基因后，运用GO（Gene Ontology）和KEGG pathway进行功能聚类分析。④挑选部分差异表达基因进行半定量PCR验证。结果 ①KCTD15基因敲低后筛选到833个差异表达基因（差异倍数值≥2或≤0.5）。②GO功能分析显示差异表达基因主要富集在以下相关通路：细胞黏附（34个），细胞骨架组装（26个）、生物膜组装（26个）、染色体组装（20个），脂肪细胞分化（6个）；KEGG pathway分析则显示半乳糖代谢和二羧酸代谢受到影响。③对MTHFD1、MDH1、VSP4B、CUX1、ABCA7 5个基因表达的半定量PCR与芯片测试结果相符。结论 KCTD15基因功能与生物膜和细胞骨架有关且能够影响细胞能量代谢过程。 [关键词] KCTD15 基因；3T3-L1前体脂肪细胞；基因芯片；肥胖 Effect of KCTD15 gene on 3T3-L1 preadipocyte by global expression profile microarray Zhang Peng1, Jin Kangxuan2, Chen Yusheng2, Xu Zihui1, 3 (1Department of Endocrinology, 3Department of Integrated Traditional and Western Medicine, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037; 2Laboratory of Major Disease Genomics and Personalized Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100029, China) [Abstract] Objective To determine the cellular function of 3T3-L1 preadipocytes with normal and knockdown potassium channel tetramerisation domain containing 15 (KCTD15) and analyze the different expressed genes (DEGs) to explore the potential relationship between KCTD15 and obesity. Methods RNAi technology was used to specifically decrease KCTD15 gene expression in 3T3-L1 preadipocyte. Then semi-quantitative reverse transcriptase PCR was employed to confirm the KCTD15 knockdown efficiency. The expression of genes at transcriptional level was detected by Mouse WG-6 V2.0 expression beadchip (Illumina). Based on the chip data, the DEGs were screened out according to fold change parameters. The obtained DEGs were further analyzed through 2 bio-informatic softwares, GO analyse and KEGG pathway. Several DEGs were selected for verification by semi-quan