Endocrine disruptive effects of cadmium on steroidogenesis: Human adrenocortical carcinoma cell line NCI-H295R as a cellular model for reproductive toxicity testing
Endocrine disruptive effects of cadmium on steroidogenesis: Human adrenocortical carcinoma cell line NCI-H295R as a cellular model for reproductive toxicity testing
复制标题
DOI：
10.1080/10934529.2015.987520
复制
发表时间：
2015-03-21
影响因子：
2.1
通讯作者：
Lukac, Norbert
中科院分区：
文献类型：
--
作者：
Knazicka, Zuzana;Forgacs, Zsolt;Lukac, Norbert
镉(Cd)是一种已知的内分泌干扰物，能够影响参与生殖过程调节的激素的产生。本研究以人肾上腺癌细胞系NCI-H295R为体外生物学模型，研究了氯化镉对类固醇合成的影响。将细胞培养物暴露于不同浓度的氯化镉(1.90、3.90、7.80、15.60、31.20和62.50 mM)中，并与对照组(不加氯化镉)进行比较。用代谢活性(MTT法)测定细胞活力，以评估线粒体结构的完整性。用酶联免疫吸附试验(ELISA法)直接定量测定培养基中性类固醇的产生。细胞在氯化镉存在下培养48h后，观察到低浓度的氯化镉对孕酮的产生有浓度依赖性的耗竭作用。在较高剂量(31.20mU/L)的氯化镉处理组中，孕酮含量最低，并具有显著的细胞毒作用(P<；0.01)。在整个应用范围内，即使在较低浓度的氯化镉下，镉也会降低睾酮的释放。17β-雌二醇的释放也有所下降，但与孕酮和睾酮的下降相比，下降的幅度不那么明显。在本研究所用的所有浓度的氯化镉(1.90~62.50 mM)下，细胞毒作用均显著(P<；0.01)。然而，细胞存活率在7.80 mU/M时仍保持较高的水平(P>；75%)，在15.60 mU/M和较高浓度时细胞存活率显著下降(P<；0.01)。这些结果表明，即使在很低的浓度下，镉对性激素的合成也有内分泌干扰作用。
Cadmium (Cd) is a known endocrine disruptor with the ability to affect the production of hormones involved in the regulation of reproductive processes. In this study human adrenocortical carcinoma cell line NCI-H295R was used as an in vitro biological model to study the effect of cadmium (CdCl2) on steroidogenesis. The cell cultures were exposed to different concentrations of CdCl2 (1.90, 3.90, 7.80, 15.60, 31.20 and 62.50 mu M) and compared to control (medium without CdCl2). Cell viability was measured by the metabolic activity (MTT) assay for estimation of mitochondria structural integrity. Quantification of sexual steroid production directly from aliquots of the medium was performed by enzyme linked immunosorbent assay (ELISA). Following 48h culture of the cells in the presence of CdCl2 a concentration-dependent depletion in progesterone production was observed at the lower concentrations of CdCl2. The lowest amount of progesterone was significantly detected in groups with the higher doses (>= 31.20 mu M) of CdCl2, which elicited significant (P < 0.01) cytotoxic action, too. Cadmium decreased testosterone release in the whole applied range even at the lower concentration of CdCl2. The release of 17 beta-estradiol decreased as well, but the decline was less pronounced compared to decrease of progesterone and testosterone. The cytotoxic effect was significantly (P < 0.01) detected at all concentrations of CdCl2 (1.90-62.50 mu M) used in the study. However, the cell viability remained relatively high (>75%) up to 7.80 mu M of CdCl2 and significantly (P < 0.01) decreased at 15.60 mu M and higher concentrations of CdCl2. These results suggest that cadmium has endocrine disruptive effects on sexual steroid synthesis even at very low concentrations.