miRNA regulation of cytotoxic effects in mouse Sertoli cells exposed to nonylphenol

Abstract

BackgroundIt is known that some environmental chemicals affect the human endocrine system. The harmful effects of endocrine disrupting chemical (EDC) nonylphenol (NP) have been studied since the 1980s. It is known that NP adversely affects physiological functions by mimicking the natural hormone 17 beta-estradiol. In the present study, we analyzed the expression of miRNAs and their target genes in mouse Sertoli TM4 cells to better understand the regulatory roles of miRNAs on Sertoli cells after NP exposure.MethodsMouse TM4 Sertoli cells were treated with NP for 3 or 24 h, and global gene and miRNA expression were analyzed using Agilent mouse whole genome and mouse miRNA v13 arrays.ResultsWe identified genes that were > 2-fold differentially expressed in NP-treated cells and control cells (P < 0.05) and analyzed their functions through Gene Ontology analysis. We also identified miRNAs that were differentially expressed in NP-treated and control cells. Of the 186 miRNAs the expression of which differed between NP-treated and control cells, 59 and 147 miRNAs exhibited 1.3-fold increased or decreased expression at 3 and 24 h, respectively. Network analysis of deregulated miRNAs suggested that Ppara may regulate the expression of certain miRNAs, including miR-378, miR-125a-3p miR-20a, miR-203, and miR-101a, after exposure to NP. Additionally, comprehensive analysis of predicted target genes for miRNAs showed that the expression of genes with roles in cell proliferation, the cell cycle, and cell death were regulated by miRNA in NP-treated TM4 cells. Levels of expression of the miRNAs miR-135a* and miR-199a-5p were validated by qRT-PCR. Finally, miR-135a* target gene analysis suggests that the generation of reactive oxygen species (ROS) following exposure to NP exposure may be mediated by miR-135a* through regulation of the Wnt/beta-catenin signaling pathway.ConclusionsCollectively, these data help to determine NP's actions on mouse TM4 Sertoli cells and increase our understanding of the molecular mechanisms underlying the adverse effects of xenoestrogens on the reproductive system.

Roles of miRNAs in microcystin-LR-induced Sertoli cell toxicity

“…The miRNA microarray results indicated that 8 miRNAs (miR-374b-5p, miR-362-5p, miR450a-5p, miR-193a-3p, miR-128-3p, miR-29b-3p, miR-199a-5p, and miR-miR-181a) involved in both azoospermia and non-obstructive azoospermia were dysregulated in MC-LR-treated Sertoli cells. miR-374b-5p, miR-128a-3p, miR-29b-3p and miR-199a-5p have been reported to be altered in nonylphenol-induced Sertoli cell toxicity (Choi et al, 2011). The dysregulation of miR-374b-5p, miR-193a-3p, miR-128-3p and miR-181a was also found in the semen of patients with azoospermia and asthenozoospermia (Wang et al, 2011).…”

“…Computational predictions indicate that more than one third of all human genes may be the targets of miRNA (Krek et al, 2005). Recent research has demonstrated that gene expression of Sertoli cells is regulated by sophisticated mechanisms including epigenomic regulation by miRNAs (Choi et al, 2011). Accumulating evidence has proven a crucial role of miRNAs in MC-induced cancer progression and embryonic development (Xu et al, 2012;Zhao et al, 2012).…”

“…We finally chose 500 nM for the subsequent array-based miRNA and mRNA profiling. We chose 2-fold for miRNA and 1.2-fold for mRNA as the cut-off as generally suggested in the microarray analysis (Choi et al, 2011;Martinez-Pacheco et al, 2014).…”

Environmental endocrine disruptors: Effects on the human male reproductive system

“…In particular, microRNAs (miR) are involved in proper differentiation of primordial germ cells (PGCs) and have been shown to be dysregulated in testicular germ cell tumors [32]. Alterations in miR expression have been observed in mouse Sertoli cells that were exposed to nonyl-phenol [33]. Other EDCs, such as BPA and DDT have also been linked to altered miR expression in estrogen-responsive human breast cancer MCF-7 cells and in placental cell lines, respectively [32].…”

Sexually dimorphic effects of gestational endocrine-disrupting chemicals on microRNA expression in the developing rat hypothalamus

“…The effects of prenatal EDCs on developmental expression of microRNAs have not been studied in the hypothalamus, but their effects have been shown in other tissues including mouse Sertoli cells (22), whole brains, and livers (23); rat penile shafts (24) and hippocampal cultures (25); and human breast carcinoma (26, 27) and placental cell lines (28). In the current study, we addressed whether a class of prenatal EDCs affect the expression of microRNAs during brain sexual differentiation following prenatal exposure of rats to A1221, a mixture of polychlorinated biphenyls (PCBs) that has previously been shown to perturb expression of sexually dimorphic genes in the brain and cause reproductive and behavioral phenotypic changes in adulthood (4, 5, 29–32).…”