Department of Urology, Children's Hospital of Chongqing Medical University, Chongqing, 400014, PR China; Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Chongqing, 400014, PR China; Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing, 400014, PR China; National Clinical Research Center for Child Health and Disorders, Chongqing, 400014, China; China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing, 400014, PR China; Chongqing Key Laboratory of Pediatrics, Chongqing, 400014, PR China.
Chongqing Key Laboratory of Children Urogenital Development and Tissue Engineering, Chongqing, 400014, PR China; Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing, 400014, PR China; National Clinical Research Center for Child Health and Disorders, Chongqing, 400014, China; China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing, 400014, PR China; Chongqing Key Laboratory of Pediatrics, Chongqing, 400014, PR China.
Environ Pollut. 2021 Jan 1;268(Pt A):115627. doi: 10.1016/j.envpol.2020.115627. Epub 2020 Sep 24.
N-methyladenosine (m6A) modification, the most prevalent form of RNA methylation, modulates gene expression post-transcriptionally. Di-(2-ethylhexyl) phthalate (DEHP) is a common environmental endocrine disrupting chemical that induces testicular injury due to the inhibition of the demethylase fat mass and obesity-associated protein (FTO) and increases the m6A modification. How FTO-mediated m6A modification in testicular Leydig cell injury induced by DEHP remains unclear. Here, the TM3 Leydig cell line was treated with mono-(2-ethylhexyl) phthalate (MEHP), the main metabolite of DEHP in the body, as well as FB23-2, an inhibitor of FTO. Decreased levels of testosterone in the culture supernatant, significantly increased apoptosis, and a remarkable upregulation of global m6A modification were found in both TM3 cells treated with MEHP and FB23-2. Transcriptome sequencing showed that both treatments significantly induced apoptosis-associated gene expression. Methylated RNA immunoprecipitation sequencing showed that the Leydig cell injury induced by upregulated m6A modification could be associated with multiple physiological disorders, including histone acetylation, reactive oxygen species biosynthesis, MAPK signaling pathway, hormone secretion regulation, autophagy regulation, and male gonadal development. Overall, the inhibition of FTO-mediated up-regulation of m6A could be involved in MEHP-induced Leydig cell apoptosis.
N6-甲基腺苷（m6A）修饰是 RNA 甲基化中最普遍的形式，可在后转录水平调节基因表达。邻苯二甲酸二（2-乙基己基）酯（DEHP）是一种常见的环境内分泌干扰化学物质，由于抑制去甲基酶脂肪量和肥胖相关蛋白（FTO），会导致睾丸损伤，并增加 m6A 修饰。DEHP 诱导的睾丸间质细胞损伤中 FTO 介导的 m6A 修饰如何发生尚不清楚。在这里，TM3 间质细胞系用 MEHP（DEHP 体内的主要代谢物）和 FB23-2（FTO 抑制剂）处理。MEHP 和 FB23-2 处理的 TM3 细胞中，培养上清液中睾酮水平降低，细胞凋亡明显增加，整体 m6A 修饰显著上调。转录组测序显示，两种处理均显著诱导凋亡相关基因表达。甲基化 RNA 免疫沉淀测序表明，上调的 m6A 修饰引起的间质细胞损伤可能与多种生理紊乱有关，包括组蛋白乙酰化、活性氧生物合成、MAPK 信号通路、激素分泌调节、自噬调节和男性性腺发育。总体而言，FTO 介导的 m6A 上调抑制可能参与 MEHP 诱导的间质细胞凋亡。
