Endocrine disruptive effects of cadmium on steroidogenesis: Human adrenocortical carcinoma cell line NCI-H295R as a cellular model for reproductive toxicity testing

Abstract

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 μM) 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 48 h 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 μM) 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 17β-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 μM) used in the study. However, the cell viability remained relatively high (>75%) up to 7.80 μM of CdCl₂ and significantly (P < 0.01) decreased at 15.60 μM and higher concentrations of CdCl₂. These results suggest that cadmium has endocrine disruptive effects on sexual steroid synthesis even at very low concentrations.

Keywords:

• Cadmium chloride
• cell viability
• endocrine disruption
• NCI-H295R cell line steroid hormones

Funding

This work was supported by the Scientific Agency of the Slovak Republic VEGA No. 1/0857/14, by European Community under project no 26220220180: Building Research Centre, AgroBioTech and the Tatra Bank Foundation 2012/2013.