Reproductive Toxicology ( IF 3.3 ) Pub Date : 2022-05-17 , DOI: 10.1016/j.reprotox.2022.05.006 Daniele Marcoccia 1 , Antonella Smeriglio 2 , Alberto Mantovani 1 , Domenico Trombetta 2 , Stefano Lorenzetti 1
Endocrine disruption mechanisms in prostate are an overlooked issue. The anti-androgenic properties of the fungicide vinclozolin (VIN) and its active metabolites - 2-[[(3,5- dichlorophenyl)-carbamoyl]oxy]− 2-methyl-3-butenoic acid (M1) and 3’5’-dichloro-2-hydroxy-2- methylbut-3-enanilide (M2) - were assessed on human prostate-derived cells (LNCaP); the effects were investigated also upon co-treatment with 5α-dihydrotestosterone (DHT), the physiological androgen receptor (AR)-agonist, and compared to the anti-androgenic drugs, 2-hydroxy-flutamide (2OH-FTA) and bicalutamide (BIC). Assessed endpoints were the cellular uptake and subcellular localization of VIN, M1 and M2, DHT-induced PSA gene expression and secretion. VIN, its metabolites, and the reference drugs, significantly reduced DHT-induced PSA secretion and gene expression, M2 showing the strongest downregulation. In absence of DHT, 2OH-FTA and BIC showed a very high (>98%) LNCaP uptake with a predominant intranuclear localization (BIC=80%, 2OH-FTA=70%). VIN cellular uptake was 42%: 24.7% made up by M2, mostly localized at nuclear level, differently from VIN and M1. Upon DHT co-treatment, VIN intracellular uptake increased by 28%, especially in the microsomal fraction (MF); M2 also increased mainly in MF but also, to a lower extent, in the intranuclear fraction. Finally, in a 72-hr time-course, the LNCaP uptake of VIN and its metabolites was much faster compared to purified M1 and M2. Overall, M2 resulted the leading compound for VIN endocrine-disrupting effects in LNCaP.
中文翻译:
长春唑啉及其代谢物的细胞内分布对 LNCaP 细胞中 5α-二氢睾酮 (DHT) 诱导的 PSA 分泌的影响不同
前列腺内分泌干扰机制是一个被忽视的问题。杀菌剂vinclozolin (VIN) 及其活性代谢物- 2-[[(3,5- dichlorophenyl)-carbamoyl]oxy]- 2-methyl-3-butenoic acid (M1) 和 3'5'的抗雄激素特性-dichloro-2-hydroxy-2-methylbut-3-enanilide (M2) - 在人类前列腺衍生细胞 (LNCaP) 上进行了评估;还研究了与 5α-二氢睾酮 (DHT)、生理性雄激素受体 (AR) 激动剂共同治疗的效果,并与抗雄激素药物 2-羟基氟他胺 (2OH-FTA) 和比卡鲁胺 (BIC) 进行比较)。评估的终点是 VIN、M1 和 M2、DHT 诱导的细胞摄取和亚细胞定位PSA基因的表达和分泌。VIN、其代谢物和参考药物显着降低了 DHT 诱导的 PSA 分泌和基因表达,M2 表现出最强的下调。在没有 DHT 的情况下,2OH-FTA 和 BIC 显示出非常高 (>98%) 的 LNCaP 摄取,主要是核内定位 (BIC=80%, 2OH-FTA=70%)。VIN 细胞摄取率为 42%:24.7% 由 M2 组成,主要位于核水平,与 VIN 和 M1 不同。在 DHT 联合处理后,VIN 细胞内摄取增加了 28%,尤其是在微粒体部分 (MF);M2 也主要在 MF 中增加,但在核内部分也有所增加。最后,在 72 小时的时间过程中,与纯化的 M1 和 M2 相比,LNCaP 对 VIN 及其代谢物的吸收要快得多。全面的,