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基于转录组测序的 GULP1 下游靶基因的筛选与分析。

Screening and analysis of GULP1 downstream target genes based on transcriptomic sequencing.

机构信息

Key laboratory of Xinjiang Endemic and Ethnic Diseases, School of Medicine,Shihezi University, Shihezi 832000, China.

出版信息

Yi Chuan. 2024 Oct;46(10):860-870. doi: 10.16288/j.yczz.24-221.

Abstract

GULP1 is an engulfment adaptor protein containing a phosphotyrosine-binding (PTB) domain, and existing studies have shown that it can promote glucose uptake in 3T3-L1 adipocytes. To further explore key metabolically related differential genes downstream of GULP1, this study conducted transcriptome analysis on adipocytes and skeletal muscle cells overexpressing GULP1. Subsequently, abnormally expressed genes were subjected to bioinformatic analysis, and real-time fluorescent quantitative PCR (qRT-PCR) was used for mutual validation with transcriptome sequencing. The results indicated that, with a threshold of < 0.05 and |LogFoldChange| ≥ 1 for screening differentially expressed genes, compared with control cells, there were 278 upregulated and 263 downregulated genes in adipocytes overexpressing GULP1. Metabolism-related GO (Gene Ontology) terms included cholesterol biosynthetic process, cholesterol metabolic process, response to lipopolysaccharide, lipid metabolic process, etc. A total of 52 metabolically related differentially expressed genes were enriched in 10 KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways, with lipid metabolism being highly enriched. In skeletal muscle cells overexpressing GULP1, there were 280 upregulated and 302 downregulated genes, with metabolism-related GO terms including hormone metabolic process, response to lipopolysaccharide, one-carbon metabolic process, etc. A total of 86 metabolically related differentially expressed genes were enriched in 10 KEGG pathways, with amino acid metabolism, lipid metabolism, and carbohydrate metabolism being highly enriched. GULP1's biological functions are extensive, including lipid metabolism and oncology. This study, through transcriptomics and bioinformatic analysis, identified key metabolically related differential genes downstream of GULP1, obtained metabolically related differential genes and signaling pathways after GULP1 overexpression, providing important theoretical basis for future research on GULP1 downstream target genes.

摘要

GULP1 是一种含有磷酸酪氨酸结合(PTB)结构域的吞噬作用衔接蛋白,已有研究表明其可促进 3T3-L1 脂肪细胞的葡萄糖摄取。为了进一步探索 GULP1 下游与代谢相关的关键差异基因,本研究对过表达 GULP1 的脂肪细胞和骨骼肌细胞进行了转录组分析。随后,对异常表达的基因进行生物信息学分析,并通过实时荧光定量 PCR(qRT-PCR)与转录组测序进行相互验证。结果表明,与对照细胞相比,过表达 GULP1 的脂肪细胞中,筛选出差异表达基因的阈值为 < 0.05 和 |LogFoldChange| ≥ 1,有 278 个基因上调,263 个基因下调。与代谢相关的 GO(基因本体论)术语包括胆固醇生物合成过程、胆固醇代谢过程、脂多糖反应、脂质代谢过程等。在 10 个 KEGG(京都基因与基因组百科全书)途径中,共富集了 52 个与代谢相关的差异表达基因,其中脂质代谢途径高度富集。在过表达 GULP1 的骨骼肌细胞中,有 280 个基因上调,302 个基因下调,与代谢相关的 GO 术语包括激素代谢过程、脂多糖反应、一碳代谢过程等。在 10 个 KEGG 途径中,共富集了 86 个与代谢相关的差异表达基因,其中氨基酸代谢、脂质代谢和碳水化合物代谢途径高度富集。GULP1 的生物学功能广泛,包括脂质代谢和肿瘤学。本研究通过转录组学和生物信息学分析,鉴定了 GULP1 下游与代谢相关的关键差异基因,获得了 GULP1 过表达后与代谢相关的差异基因和信号通路,为未来研究 GULP1 下游靶基因提供了重要的理论依据。

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