Identification of genes showing altered DNA methylation and gene expression in the renal proximal tubular cells of rats treated with ochratoxin A for 13 weeks.

Ochratoxin A (OTA) is a mycotoxin that causes renal carcinogenicity following the induction of karyomegaly in proximal tubular cells after repeated administration to rats. Here, we performed gene profiling regarding altered DNA methylation and gene expression in the renal tubules focusing on the mechanism of OTA‐induced carcinogenesis. For this purpose, OTA or 3‐chloro‐1,2‐propanediol (3‐MCPD), a renal carcinogen not inducing karyomegaly, was administered to rats for 13 weeks, and DNA methylation array and RNA sequencing analyses were performed on proximal tubular cells. Genes for which OTA altered the methylation status and gene expression level, after excluding genes showing similar expression changes by 3‐MCPD, were subjected to confirmation analysis of the transcript level by real‐time reverse‐transcription PCR. Gene Ontology (GO)‐based functional annotation analysis of validated genes revealed a cluster of hypermethylated and downregulated genes enriched under the GO term "mitochondrion," such as those associated with metabolic reprogramming in carcinogenic process (Clpx, Mrpl54, Mrps34, and Slc25a23). GO terms enriched for hypomethylated and upregulated genes included "response to arsenic‐containing substance," represented by Cdkn1a involved in cell cycle arrest, and "positive regulation of IL‐17 production," represented by Osm potentiating cell proliferation promotion. Other genes that did not cluster under any GO term included Lrrc14 involved in NF‐κB‐mediated inflammation, Gen1 linked to DNA repair, Has1 related to chromosomal aberration, and Anxa3 involved in tumor development and progression. In conclusion, a variety of genes engaged in carcinogenic processes were obtained by epigenetic gene profiling in rat renal tubular cells specific to OTA treatment for 13 weeks. The present study investigated comprehensive gene profiles regarding DNA methylation and gene expression in rat renal tubules after 13‐week administration of ochratoxin A (OTA), a karyomegaly‐inducing renal carcinogen. OTA‐specific genes were obtained by excluding those responding to 3‐MCPD, a renal carcinogen not inducing karyomegaly. Genes obtained included those associated with metabolic reprogramming, cell cycle arrest, cell proliferation facilitation, inflammation, DNA repair, chromosomal aberration, and tumor development and progression, suggesting that various gene functions are epigenetically engaged in OTA‐specific carcinogenic processes. [ABSTRACT FROM AUTHOR]