Gene expression analysis of toxicological pathways in TM3 leydig cell lines treated with Ethane dimethanesulfonate

Abstract

Ethane dimethanesulfonate (EDS), a well-known alkylating agent, selectively destroys Leydig cells. To clarify the molecular pathways underlying EDS action on Leydig cells, we analyzed gene expression profiles of an EDS-treated TM3 Leydig cell line. In this study, we analyzed the representative canonical pathways and toxicity pathways/gene lists using the Ingenuity Pathways Analysis program. In TM3 cells, 677 and 6756 genes were identified as being up- or downregulated after 3 and 24 h EDS treatments, respectively, (>1.3-fold changes, p < 0.05). Toxicological pathway analysis revealed that expression of genes related to Nrf2-mediated oxidative stress response showed remarkable changes in early or later stage of EDS-treated TM3 cells. Several genes related to steroidogenesis and apoptosis were also differentially expressed at 24 h in EDS-treated TM3 cells. Overall, toxicological pathway analysis using gene expression profiling showed that oxidative stress might be an important factor in cell death in TM3 cells affected by EDS treatment.

Identification of Stem Leydig Cells Derived from Pig Testicular Interstitium

“…The EDS was provided by Professor Yuanqiang Zhang (Department of Human Anatomy, Histology and Embryology, The Fourth Military Medical University, China). According to the previous methods, EDS was dissolved in dimethyl sulfoxide (DMSO)/sterile water (1 : 3, v/v) [ 23 – 25 ]. Afterwards, the primary isolated SLCs were seeded into a 6-well plate and 0, 0.5, 0.75, and 1.0 mg/mL EDS (final concentration) were added to the culture solution, respectively [ 24 , 25 ].…”

miR-205 Expression Elevated With EDS Treatment and Induced Leydig Cell Apoptosis by Targeting RAP2B via the PI3K/AKT Signaling Pathway

“…In this study, EDS could eliminate differentiated LCs in pigs, which was accompanied with a decreased expression of CYP17A1, whereas the expression of PDGFRα did not change. The molecular pathway mechanisms underlying EDS action on LCs were related to intracellular glutathione (Kelce and Zirkin, 1993), activation of the Fas receptor (Taylor et al, 1999), oxidative stress (Lee et al, 2012), involvement of Bcl-2 family members (Taylor et al, 1998), or alkylation of proteins (Rommerts et al, 1985). In this study, we first uncovered that miR-205 was also involved in the process of EDS to eliminate differentiated LCs.…”

Ethylene dimethanesulfonate effects on gene promoter activities related to the endocrine function of immortalized Leydig cell lines R2C and MA-10

“…In addition to being efficient in whole animal models, in vitro approaches using EDS on different Leydig cell lines have revealed that EDS is similarly effective as in vivo approaches. Although cell death was triggered in EDS-treated rat (H540) and mouse (MA-10 and TM3) Leydig cell lines, the differential sensitivity to EDS was maintained, with rat Leydig cell lines being more sensitive to EDS than the mouse cell lines ( Rommerts et al, 2004 , King et al, 1998 , Lee et al, 2012 , Li et al, 1822 ). These studies also highlighted some of the pathways that are affected by EDS treatment causing Leydig cell death.…”

“…As revealed by in vivo approaches and studies using Leydig cell lines, EDS invariably leads to Leydig cell death mainly by apoptosis involving the caspase signaling pathway ( Teerds and Rijntjes, 2007 , Rommerts et al, 2004 , King et al, 1998 ). There is also evidence that oxidative stress might trigger these events leading to apoptosis ( Lee et al, 2012 ). However, before apoptosis is triggered, there is a significant decrease in steroidogenesis and loss of steroidogenic function.…”

“…However, before apoptosis is triggered, there is a significant decrease in steroidogenesis and loss of steroidogenic function. This is mainly attributed to an important reduction in the expression of the Star gene, which codes for the steroidogenic acute regulatory protein (STAR), a key protein for steroidogenesis ( Teerds and Rijntjes, 2007 , King et al, 1998 , Lee et al, 2012 ). In addition, EDS was found to activate the transcription factor NF-κB, which binds to and activates the Ndrg2 promoter leading to increased levels of NDRG2, a pro-apoptotic protein ( Li et al, 1822 ).…”