The Role of miRNAs in Zearalenone-Promotion of TM3 Cell Proliferation

Abstract

Zearalenone (ZEA) is a non-steroidal estrogen mycotoxin produced by several Gibberella and Fusarium species. Accumulating evidence has indicated that ZEA strongly stimulates cell proliferation. However the detailed molecular and cellular mechanisms of ZEA-mediated induction of cell proliferation have not yet been completely explained. The aim of this study was to detect the role of miRNAs in ZEA-mediated induction of cell proliferation. The effects of ZEA on cell proliferation were assessed using a cell counting kit assay and xCELLigence system. Micro-RNA sequencing was performed after treatment of TM3 cells with ZEA (0.01 μmol/L) for different time periods (0, 2, 6 and 18 h). Cell function and pathway analysis of the miRNA target genes were performed by Ingenuity Pathway Analysis (IPA). We found that ZEA promotes TM3 cell proliferation at low concentrations. miRNA sequenceing revealed 66 differentially expressed miRNAs in ZEA-treated cells in comparison to the untreated control ( p < 0.05). The miRNA sequencing indicated that compared to control group, there were 66 miRNAs significant change (p < 0.05) in ZEA-treated groups. IPA analysis showed that the predicated miRNAs target gene involved in cell Bio-functions including cell cycle, growth and proliferation, and in signaling pathways including MAPK and RAS-RAF-MEK-ERK pathways. Results from flow cytometry and Western Blot analysis validated the predictions that ZEA can affect cell cycle, and the MAPK signaling pathway. Taking these together, the cell proliferation induced ZEA is regulated by miRNAs. The results shed light on the molecular and cellular mechanisms for the mediation of ZEA to induce proliferation.

Zearalenone and the Immune Response

“…Similar results on Leydig cells were found by Wang et al [ 101 ], with ZEA 50 μM. By contrast, Zheng et al [ 89 ], found that low doses of zearalenone (0.01, 0.02, 0.03, 0.04, and 0.05 μmol/L) stimulated cell viability of TM3 cells (Leydig cells) measured by using the xCELLigence real-time cell analysis. Also, under the action of ZEA (20μM), cell viability of Sertoli cells derived from Male Wistar, which are important for male reproductive system, increased over control [ 102 ].…”

“…The study of Zhang et al [ 88 ] demonstrated that ZEA at 10–50 μg/mL had a time and dose dependent inhibitory action on mouse thymic epithelial cells proliferation and arrested thymic cells in G2/M phase of cellular cycle. Studies performed on TM3 cells shows that low doses of ZEA increases cell proliferation [ 89 ].…”

Cytoprotective, Antiproliferative, and Anti-Oxidant Potential of the Hydroethanolic Extract of Fridericia chica Leaves on Human Cancer Cell Lines Exposed to α- and β-Zearalenol

“…The finding in terms of proliferation reported here may be due to the expression of two ER (α-ER and β-ER), which has also been reported for other xenoestrogens acting on liver cancer cells [ 42 ]; however, signaling pathways are not yet fully established. It is plausible that micro-RNAs are also involved in stimulating cell proliferation by silencing various cell cycle inhibitors; accordingly, it has been stated that ZEN induces oxidative stress, which may be a regulator of miRNAs, interfering with micro-RNA expression [ 43 ].…”

Toxicodynamic of combined mycotoxins: MicroRNAs and acute‐phase proteins as diagnostic biomarkers

“…The toxicodynamic effects of ZEA, a mycotoxin produced by Fusarium species, involve complex molecular interactions, and emerging research has highlighted the role of regulatory miRNAs in orchestrating the cellular response to ZEA exposure. Study has identified specific miRNAs, including miR‐22 and miR‐155, as key modulators of pathways associated with ZEA toxicity (Zheng et al., 2019). These miRNAs are implicated in regulating cellular processes such as apoptosis, inflammation, and hormonal signaling, which are perturbed by ZEA.…”