MicroRNA-375 Mediated Regulation on Pre-mRNA Processing Factor 3 in Zebrafish Embryos Exposed to Di-(2-ethylhexyl)phthalate at Low Concentrations

Di-(2-ethylhexyl)phthalate (DEHP) is an endocrine-disrupting chemical (EDC) that induces epigenetic alterations, apoptosis, and oxidative stress after biological exposure. MicroRNAs (miRNAs) are a class of small noncoding RNAs with many regulatory functions and play a role in organisms exposed to environmental chemicals. miRNA-mRNA prediction indicated that pre-mRNA processing factor 3 (PRPF3) is a likely target mRNA for miR-375 whose expression is altered by DEHP exposure. However, the interrelation between miR-375 and PRPF3 has not yet been confirmed experimentally. This study aimed to investigate the effects of DEHP on miR-375 and PRPF3 in zebrafish. The expression of miR-375 was downregulated, whereas PRPF3 was upregulated at both transcriptional and post-transcriptional levels upon stimulation with DEHP. The interaction between miR-375 and the 3′-untranslated region (3′-UTR) of PRPF3 was confirmed by a dual fluorescent protein assay and a dual luciferase reporter gene assay. The expression of PRPF3 at both transcriptional and post-transcriptional levels was reduced in ZF4 cells when transfected with a miR-375 mimic but increased when transfected with a miR-375 inhibitor. The results improved our understanding of molecular mechanisms of toxicity upon DEHP exposure and presented miR-375 as a potential novel toxicological biomarker for chemical exposure.