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.. Figure 3 Expression pattern of androgen-synthesizing enzymes in normal adrenals and the androgen-secreting oncocytoma. (A) Expression mRNA levels of CYB5, AKR1C3 and HSD17B3 normalized to PPIA (cyclophilin) in normal adrenals (white bars) and two samples of the ... Table 3 Scores of steroidogenic enzyme immunoreactivities in normal and tumor adrenocortical tissues. Among transcripts encoding the different isoforms of 17��-HSD, which are enzymes catalyzing the conversion of ��4-androstenedione into testosterone, AKR1C3 mRNA (encoding Oxygenase 17��-HSD5) levels were higher than those of HSD17B3 (encoding 17��-HSD3) in both the adenoma and normal adrenal tissues (Fig. 3A). Both 17��-HSD5 and 17��-HSD3 immunoreactivities were observed in zona glomerulosa IOX1 chemical structure and ZR of normal adrenals (Fig. 3B). In the adrenocortical lesion, weak labeling for 17��-HSD5 and intense 17��-HSD3 immunostaining were detected in clusters of oncocytic cells (Fig. 3C and Table 3). Expression of CYP19A1 encoding aromatase, the estrogen synthesizing enzyme, was low and variable in normal adrenal samples and was negligible in the adenoma tissue (not shown). Steroid hormone secretion Cultured adrenocortical cells derived from the adenoma secreted for 24?h significant amounts of ��4-androstenedione (4.1��0.3?ng/106 cells), testosterone (2.3��0.2?ng/106 cells) and cortisol (24.3��2.5?ng/106 cells) (Fig. 4A). Incubation of cells with increasing concentrations of ACTH (10?12�C10?9?M) provoked dose-dependent stimulations of ��4-androstenedione (+68.1��3.8%), testosterone (+40.6��4.4%) and cortisol (+181.3��8.9%) secretions (Fig. 4B, C and D). ACTH was more efficient for stimulating ��4-androstenedione (P=0.003) and cortisol (PCapmatinib manufacturer of ACTH on cortisol secretion was weak in tumor cells in comparison with normal adrenocortical cells (approximately+1000%) (23). The Leydig-cell-like expression pattern of steroidogenic enzymes prompted us to investigate the involvement of LH/hCG receptors in the control of steroidogenesis in adenoma cells. hCG (10?12�C10?7?M) had no effect on steroid production. Figure 4 Steroid secretion by cultured adrenocortical oncocytoma cells derived from the patient with hirsutism. (A) Spontaneous secretions of ��4-androstenedione, testosterone and cortisol by cultured oncocytoma cells. (B, C and D) Effects of increasing ... Expression of ACTH receptor Expression level of MC2R mRNAs, encoding the MC2R selective for ACTH, in the tissue of the patient was four times lower than that detected in normal adrenals (Fig. 4E). Intense MC2R immunoreactivity was observed throughout the ZF of normal adrenals whereas weak labeling was present in clusters of oncocytic cells in the adenoma (Fig. 4F). As expected, no LH receptor-immunostaining was detected in the adenoma tissue (not shown).
