MEHP activates JNK to inhibit the migration of human foreskin fibroblasts

Abstract

This study aimed to investigate the impact of mono(2-ethylhexyl) phthalate (MEHP) on the proliferation, apoptosis, and migration of human foreskin fibroblast cells (HFF-1) and the role of the JNK signaling pathway in cell migration. HFF-1 cells were randomly assigned to the control group with 0 MEHP exposure (M0) or the experimental groups with 25, 50, 100, 200, and 400 μmol/L MEHP exposure (M25, M50, M100, M200, and M400, respectively). After 24 and 48 h of MEHP exposure, the proliferation of HFF-1 cells in any group had no significant change. However, compared with the M0 group, the M200 and M400 groups presented substantially increased apoptosis of HFF-1 cells. Moreover, cell migration ability significantly decreased in all groups (p < 0.05). Additionally, the transcription and phosphorylated protein activation of JNK kinase in HFF-1 cells were substantially upregulated with the increase in MEHP exposure. Subsequently, HFF-1 cells were randomly divided into three groups: the DMSO blank control group, the 100 μM MEHP experimental group (M100), and the 100 μM MEHP plus 10 μM SP600125 (specific JNK inhibitor) experimental group (S10). The activation of JNK protein in HFF-1 cells was substantially downregulated in the S10 group. HFF-1 cells were also divided into the blank control group (M0). They were treated with 100 μM MEHP and varying concentrations of SP600125 (5, 10, and 15 μM for S5, S10, and S15, respectively). As the concentration of the antagonist increased, the migration ability of HFF-1 cells was returned to normal. Finally, the ROS in HFF-1 cells increased under MEHP exposure. This finding indicates that the regulation of cell migration by the JNK signaling pathway may be important in the occurrence of hypospadias.

Keywords:

• MEHP
• HFF-1
• JNK
• cell migration

Ethics approval

All research reported in submitted papers has been conducted in an ethical and responsible manner, and is in full compliance with all relevant codes of experimentation and legislation.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Authors’ contributions

Contributed to study conception and design: HZ, XY, GQZ, MYL; performed the experiments: HZ, XY, GQZ, MYL; analyzed the data: HZ, WH; contributed materials and facilities: XWF, WJ, LQ; drafted the manuscript: MYL, HZ, XY, GQZ and WH. All authors approved any revisions and the final paper.

Additional information

Funding

This study was supported by the Natural Science Foundation of Hunan Province (2020JJ4542), the Clinical Research Project of University of South China (USCKF201902K01), and the Hunan Provincial Clinical Medical Technology Innovation Guiding Project (2020SK51801).