Human adrenal corticocarcinoma NCI-H295R cells produce more androgens than NCI-H295A cells and differ in 3β-hydroxysteroid dehydrogenase type 2 and 17,20 lyase activities

Abstract

The human adrenal cortex produces mineralocorticoids, glucocorticoids, and androgens in a species-specific, hormonally regulated, zone-specific, and developmentally characteristic fashion. Most molecular studies of adrenal steroidogenesis use human adrenocortical NCI-H295A and NCI-H295R cells as a model because appropriate animal models do not exist. NCI-H295A and NCI-H295R cells originate from the same adrenocortical carcinoma which produced predominantly androgens but also smaller amounts of mineralocorticoids and glucocorticoids. Research data obtained from either NCI-H295A or NCI-H295R cells are generally compared, although for the same experiments no direct comparison between the two cell lines has been performed. Therefore, we compared the steroid profile and the expression pattern of important genes involved in steroidogenesis in both cell lines. We found that steroidogenesis differs profoundly. NCI-H295A cells produce more mineralocorticoids, whereas NCI-H295R cells produce more androgens. Expression of the 3b-hydroxysteroid dehydrogenase (HSD3B2), cytochrome b5, and sulfonyltransferase genes is higher in NCI-H295A cells, whereas expression of the cytochrome P450c17 (CYP17), 21-hydroxylase (CYP21), and P450 oxidoreductase genes does not differ between the cell lines. We found lower 3b-hydroxysteroid dehydrogenase type 2 but higher 17,20-lyase activity in NCI-H295R cells explaining the 'androgenic' steroid profile for these cells and resembling the zona reticularis of the human adrenal cortex. Both cell lines were found to express the ACTH receptor at low levels consistent with low stimulation by ACTH. By contrast, both cell lines were readily stimulated by 8Br-cAMP. The angiotensin type 1 receptor was highly expressed in NCI-H295R than NCI-H295A cells and angiotensin II stimulated steroidogenesis in NCI-H295R but not NCI-H295A cells. Our data suggest that comparative studies between NCI-H295A and NCI-H295R cells may help find important regulators of mineralocorticoid or androgen biosynthesis.

Leukemia inhibitory factor as a regulator of steroidogenesis in human NCI-H295R adrenocortical cells

“…Careful time-dependent analyses were not possible for aldosterone production due to its low production rate and relative insensitivity of the RIA. In any case, LIF also increased aldosterone production by 48-h treatment in our NCI-H295R cells, although this cell line produces less mineralocorticoids than the NCI-H295A variant (Samandari et al 2007). Our time-course experiments showed that the stimulatory effects of LIF and 8-Br-cAMP on cortisol production were somewhat slower than on adrenal androgen production.…”

“…It is noteworthy that there are two variants (NCI-H295A and NCI-H295R) of the original NCI-H295 cell line. These variants have somewhat different steroidogenic secretory profiles (Samandari et al 2007). We suppose that Bamberger et al (2000) used the parent cell line NCI-H295 in their LIF studies, although the source of their cells was not clearly described.…”

IGF2 and IGF1R in pediatric adrenocortical tumors: roles in metastasis and steroidogenesis

“…In agreement, we found significant positive correlation between IGF1R gene expression and DHEAS levels in diagnosis patient's serum. Moreover, cells treated with OSI-906 presented lower expression of genes related to hormone synthesis/metabolism and STAR protein impairment of 55%, which is considered the first key mediator of steroidogenesis (Samandari et al 2007).…”

Morphofunctional effects of mitotane on mitochondria in human adrenocortical cancer cells

“…Different strains of H295 cells have shown different degrees of inhibition in steroid secretion, depending on their steroidogenic profiles (Samandari et al 2007). In contrast to the inhibitory effect on steroid production, the cytotoxic effects of MTT responsible for adrenal necrosis are less clear.…”