show Abstracthide AbstractWe investigated the biological effects of ZEA exposure on donkey granulosa cells by using RNA-seq analysis. ZEA at 10 and 30 µM were administered to granulosa cells within 72 hours of in vitro culture. ZEA at 10 µM significantly altered the tumorigenesis associated genes in donkey granulosa cells. Exposure to 10 and 30 µM ZEA treatment significantly reduced mRNA expression of PTEN, TGFß, ATM, and CDK2 genes, particularly, the ZEA treatment significantly increased the expression of PI3K and AKT genes. Furthermore, immunofluorescence, RT-qPCR, and Western blot analysis verified the gene expression of ZEA-exposed granulosa cells. Collectively, these results demonstrated the deleterious effect of ZEA exposure on the induction of ovarian cancer related genes via the PTEN/PI3K/AKT signaling pathway in donkey granulosa cells in vitro. Overall design: 9 mRNA profiles of donkey granulosa cells were generated by RNA sequencing.