Human Adrenocortical Carcinoma (NCI-H295R) Cell Line as an <i>In Vitro</i> Cell Culture Model for Assessing the Impact of Iron on Steroidogenesis

Kňažická, Z.; Fialková, Veronika; Ďúranová, H.; Bilčíková, J.; Kováčiková, E.; Miškeje, M.; Valková, V.; Forgács, Z.; Roychoudhury, S.; Massányi, P.; Lukáč, N.

doi:10.14712/fb2021067020076

Folia Biologica > Archive > 2021, Vol. 67 > Issue 2 > Human Adrenocortical Carcinoma…

Fol. Biol. 2021, 67, 76-81

https://doi.org/10.14712/fb2021067020076

Human Adrenocortical Carcinoma (NCI-H295R) Cell Line as an In Vitro Cell Culture Model for Assessing the Impact of Iron on Steroidogenesis

Z. Kňažická¹, Veronika Fialková², H. Ďúranová², J. Bilčíková^2,3, E. Kováčiková², M. Miškeje², V. Valková², Z. Forgács⁴, S. Roychoudhury⁵, P. Massányi⁶, N. Lukáč⁶

¹Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Nitra, Slovak Republic
²AgroBioTech Research Centre, Slovak University of Agriculture, Nitra, Slovak Republic
³Department of Genetics and Plant Breeding, Faculty of Agrobiology and Food Resources, Slovak University of Agriculture, Nitra, Slovak Republic
⁴Independent Researcher, Budapest, Hungary
⁵Department of Life Science and Bioinformatics, Assam University, Silchar, India
⁶Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture, Nitra, Slovak Republic

The aim of this in vitro study was to examine the dose-dependent effects of iron as a potential endocrine disruptor in relation to the release of sexual steroid hormones by a human adrenocortical carcinoma (NCI-H295R) cell line. The cells were exposed to different concentrations (3.90, 62.50, 250, 500, 1000 μM) of FeSO₄.7H₂O and compared with the control group (culture medium without FeSO₄.7H₂O). Cell viability was measured by the metabolic activity assay. Quantification of sexual steroid production was performed by enzyme-linked immunosorbent assay. Following 48 h culture of the cells in the presence of FeSO₄.7H₂O, significantly (P < 0.001) increased production of progesterone was observed at the lowest concentration (3.90 μM) of FeSO₄.7H₂O, whereas the lowest release of progesterone by NCIH295R cells was noted after addition of 1000 μM of FeSO₄.7H₂O, which did not elicit cytotoxic action (P > 0.05). Testosterone production was substantially increased at the concentrations ≤ 62.50 μM of FeSO₄.7H₂O. Lower levels of testosterone were recorded in the groups with higher concentrations (≥ 250 μM) of FeSO₄.7H₂O (P > 0.05). The presented data suggest that iron has no endocrine disruptive effect on the release of sexual steroid hormones, but its toxicity may be reflected at other points of the steroidogenesis pathway.

Keywords

NCI-H295R cell line, ferrous sulphate heptahydrate, endocrine disruption, sexual steroid hormones, cell viability.

Funding

The study was financially supported by the Scientific Agency of the Slovak Republic VEGA 1/0163/18, APVV-16-0289, APVV-15-0543 and SUA grants 33/2019 and 35/2019.