Integration of miRNA profiling and proteomics to determine the target(s) implicated in endometrial cancer cells exposed to zearalenone.

Abstract

Zearalenone (ZEA), a natural estrogenic mycotoxin, can cause serious harmful effects on human and animal health. Several studies have reported that ZEA induces proliferation in endometrial cancer; however, its role in modulating microRNA (miRNA) and protein expression profiles remains poorly established. This study investigates the effects of low-dose ZEA exposure on endometrial cancer cells using an integrated approach combining miRNA and proteome profiling. Ishikawa cells were exposed to 25 nM ZEA or 10 nM 17-β-estradiol (E2) for 10 days. miRNA profiling using a customized cancer-specific 56-miRNA array panel revealed significant alterations in miRNA expression, with miR-204 highly upregulated in both the E2 and ZEA treatment groups. For the global protein profiling, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed. Proteomics analysis identified 243 upregulated and 227 downregulated proteins following ZEA exposure, and, in the case of E2, 286 were upregulated and 123 were downregulated. Notably, miR-204 upregulation correlated with the downregulation of three target proteins: RETSAT, IPO8, and LRRC59. Gain- and loss-of-function studies using miR-204 mimics and inhibitors confirmed its role in promoting cell proliferation, growth, and migration. The present study provides novel insights into the molecular mechanism underlying ZEA-induced promotion of endometrial cancer, highlighting the potential role of miR-204 as a key mediator. Our findings contribute to understanding the role of environmental factors in cancer development and may inform future strategies for the prevention and treatment of endometrial cancer.