Transcriptome profiling and pathway analysis of the effects of mono‑(2‑ethylhexyl) phthalate in mouse Sertoli cells

Abstract

Phthalates are confirmed to have toxic effects on the reproductive system and are likely to have further damaging actions in humans. The present study explored the molecular mechanisms of the toxic effect of mono-(2-ethylhexyl) phthalate (MEHP) on mouse Sertoli cells. Cell apoptosis and proliferation assays were used to assess the effects of MEHP on the TM4 Sertoli cell line derived from mouse testes. TM4 cells were treated with two doses of MEHP or left untreated as a control group, followed by RNA extraction and analysis using high-throughput transcriptome sequencing technology. The gene expression profile obtained was then subjected to a bioinformatics analysis to explore the molecular mechanisms of reproductive toxicity. The results revealed that 528 and 269 genes were upregulated in the high-and low-dose MEHP groups of cells compared with the control group, while 148 and 173 genes were downregulated. Gene ontology (GO) analysis indicated that the differently expressed genes were associated with the GO term 'extracellular region' of the cellular component domain in the high and low MEHP groups. Compared with the control group, eight common pathway changes were identified in the high-and low-dose MEHP groups, including 'terpenoid backbone biosynthesis'. Reverse transcription-quantitative polymerase chain reaction analysis was used to validation, and hermetic effects were observed for certain genes. These results provide an important basis and experimental data for further research into the mechanisms of phthalate-induced toxicity.

Impact of endocrine disrupting chemicals and pharmaceuticals on Sertoli cell development and functions

“…Furthermore, the sex determination pathway, Gadd45g → Gata4/Fog2 → Sry → Sox9 → FGF9 was altered in the same treatment conditions. A more recent study of MEHP exposure in mouse Sertoli cell line TM4 was performed to assess transcriptomic alterations ( 113 ). Based on high-throughput sequencing of RNA collected from TM4 cells, genes that were differentially expressed associated with the Gene Ontology (GO) term “extracellular region” when treated with high and low MEHP doses.…”

Astrocytic YAP prevents the demyelination through promoting expression of cholesterol synthesis genes in experimental autoimmune encephalomyelitis

“…The relative mRNA expression levels were represented as ΔCt = Ct gene − Ct reference, and the fold change of gene expression was calculated by using the 2 −ΔΔCt method. The primers were described previously [46][47][48][49] and were synthesized by Sangon Biotech and presented as follows: HMGCR, 5′-AGT CAG TGG GAA CTA TTG CAC-3′ and 5′-TTA CGT CAA CCA TAG CTT CCG-3′; FDPS, 5′-AGG AGG TCC TAG AGT ACA ATG CC-3′ and 5′-TGA GGG AAG AGT CCA TGA TGT C-3′; HMGCS1, 5′-TGT TCT CTT ACG GTT CTG GC-3′ and 5′-AAG TTC TCG AGT CAA GCC TTG-3′; GAPDH, 5′-AGG TCG GTG TGA ACG GAT TTG-3′ and 5′-TGT AGA CCA TGT AGT TGA GGT CA-3′.…”