A Hightroughput Approach for Screening Potential Green Plasticizers Using A Human Prostate Cell Line [set2]. Homo sapiens

The prostate is one of the main accessory glands of the male reproductive system, responsible of up to 30% of the secretions constituting seminal plasma, and is essential for male fertility. Animal studies have shown that phthalates can trigger cellular events associated with the onset of prostatic diseases. There is increasing interest in developing safe alternative plasticizers to phthalates. Many potential candidates were recently reported, and their mechanisms of action on prostatic cells warrant investigation. The goal of this study was to evaluate the toxicity of four families of chemicals that are candidate plasticizer using a non-cancerous human prostate cell line, PNT1A. These candidate plasticizers were compared to diethylhexyl phthalate (DEHP), its main bioactive metabolite monohexylethyl phthalate (MEHP), and a current “green” plasticizer that is being used increasingly, 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH), using a combination of classical (MTT) and high throughput techniques (HCS, microarray). High content screening provided additional information about cell counts and range of parameters related to cell function. The incorporation of gene expression studies revealed that several of the compounds tested in this study had an effect on cellular functions, even those that had not shown any effect with the other assays. While DINCH was found to have wide ranging effects, we show that candidate plasticizers of the dibenzoate family have no effects on metabolic activity, cellular morphology, cellular proliferation or gene expression in PNT1A cells. Candidate plasticizers of the succinate family had an effect only at the gene expression level. Overall design: Three treatments are compared to the 1% DMSO control: 3 compounds (MEHP, BDB, DOS) at 10E-4M (high). Each treatment was analyzed in quadruplicate.

前列腺是男性生殖系统的主要附属腺体之一，其分泌液可占精浆的30%，对男性生育能力至关重要。动物研究表明，邻苯二甲酸盐可诱发与前列腺疾病发作相关的细胞事件。当前，研发邻苯二甲酸盐安全替代增塑剂的关注度与日俱增。近期已有多种潜在候选物质被报道，但其对前列腺细胞的作用机制仍有待深入探究。本研究旨在利用非癌性人前列腺细胞系PNT1A，评估四类候选增塑剂的毒性。实验将这些候选增塑剂与邻苯二甲酸二(2-乙基己)酯（diethylhexyl phthalate, DEHP）、其主要活性代谢物邻苯二甲酸单己基乙酯（monohexylethyl phthalate, MEHP），以及当前日益广泛使用的“绿色”增塑剂1,2-环己烷二甲酸二异壬酯（1,2-cyclohexane dicarboxylic acid diisononyl ester, DINCH）进行对比，结合经典技术（MTT法）与高通量技术（高内涵筛选High Content Screening, HCS、基因芯片microarray）开展实验。高内涵筛选可提供关于细胞数量及多项细胞功能相关参数的额外信息。基因表达分析结果显示，本研究中测试的多种化合物会对细胞功能产生影响，即便部分化合物在其他检测方法中未表现出任何作用。尽管DINCH被发现具有广泛的细胞影响，本研究结果表明，二苯甲酸酯类候选增塑剂对PNT1A细胞的代谢活性、细胞形态、细胞增殖或基因表达均无影响。琥珀酸酯类候选增塑剂仅在基因表达层面产生影响。实验设计概述：设置三组处理组与1%二甲基亚砜（dimethyl sulfoxide, DMSO）对照组进行对比，分别为浓度10^-4 M（高浓度）的3种化合物（MEHP、BDB、DOS）。每组处理均设置四次生物学重复。