基因表达谱技术分析棒曲霉毒素肾细胞毒性的机制

Mechanisms of Patulin(PAT) Toxicity towards Human Embryonic Kidney (HEK)293 Cells, as Determined by Digital Gene Expression Profile Analysis

本文研究了食品污染物棒曲霉毒素诱导人胚肾细胞损伤过程中基因表达谱动态变化。5μM PAT作用HEK293细胞3、24h,应用数字基因表达谱(DGE)技术筛选差异表达基因,分析基因富集的GO功能,KEGG信号通路及Pathway信号途径分析,并通过定量PCR对部分差异表达基因进行验证。分析发现,在3 h和24 h受PAT诱导的差异表达基因分别为79个和261个,其中上调表达基因分别为71和192个,下调表达基因分别为8和69个。PAT主要诱导了细胞凋亡基因的差异表达,RNA降解差异基因全部表达上调,RNA合成差异基因表达下调,泛素介导的蛋白降解大部分差异基因上调,MAPK信号通路主要参与PAT毒性的下游进程,其相关差异基因大部分上调。DGE技术为我们在分子水平上提供了PAT胚肾细胞损伤机制的众多线索,为相关基因生物学效应研究奠定基础。

The dynamic changes in gene expression profiles of human embryonic kidney（HEK）293 cells during the injury process induced by food contaminant patulin（PAT） were analyzed. HEK293 cells were treated with 5 μM PAT for 3 h and 24 h, and the differentially expressed genes were screened using a digital gene expression（DGE） tag profiling technique. Gene ontology（GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway analysis were carried out, and some of the differentially expressed genes were verified by real-time quantitative polymerase chain reaction（PCR）. The results showed that 79 and 261 genes were differentially expressed after 3 h and 24 h PAT induction, respectively. Among them, 71 and 192 genes were upregulated, while 8 and 69 genes were downregulated. PAT mainly induced the differential expression of apoptosis-related genes; differentially expressed RNA degradation genes were all upregulated, and differentially expressed RNA polymerase genes were downregulated. Most differentially expressed genes of ubiquitin-mediated proteolysis were upregulated. Mitogen-activated protein kinase（MAPK） cascade signaling genes were mainly involved in the downstream process, and most related genes were upregulated. The DGE technique provided numerous clues to renal cell damage induced by PAT at the molecular level and laid a foundation for the study of the biological effects of related genes.