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纸质出版:2014
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张鹏,金康宣,陈宇晟,徐梓辉.表达谱芯片分析KCTD15对3T3-L1前体脂肪细胞的影响[J].陆军军医大学学报(原第三军医大学学报),2014,36(09):937-940.
Zhang Peng,Jin Kangxuan,Chen Yusheng,Xu Zihui.Effect of KCTD15 gene on 3T3-L1 preadipocyte by global expression profile microarray[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(09):937-940.
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RT-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Gene Ontology(GO)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RT-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RT-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。
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 the 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
Gene Ontology (GO) analysis and KEGG pathway. Several DEGs were selected for verification by semi-quantitative reverse transcriptase PCR. Results A total of 833 DEGs were identified in KCTD15 knockdown RNA sample (Fold change ≥2 or ≤0.5). GO analysis suggested that DEGs were enriched in the following pathways: biological adhesion (34)
cytoskeleton organization (26)
membrane organization (26)
chromosome organization (20) and fat cell differentiation (6). KEGG pathway analysis mapped DEGs into 2 pathway
galactose metabolism and glyoxylate and dicarboxylate metabolism. Finally
semi-quantitative reverse transcriptase PCR was used to verify the chip data on genes MTHFD1
MDH1
VSP4B
CUX1 and ABCA7. Conclusion KCTD15 gene is related to biomembrane and cytoskeleton
and affects energy metabolism process.
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