Analysis of growth factors and cytokines in human adrenocortical steroidogenesis

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We examined expression of TGF-alpha, EGF and EGFR in human adrenal cortex and its disorders, at both protein and mRNA levels. TGF-alpha expression was detected at both protein and mRNA levels in normal and neoplastic adrenals but not EGF.EGFR immunoreactivity was more widely distributed than TGF-alpha immunoreactivity, TGF-alpha and EGF was considered to be involved in biological function including adrenocortical malignancy.We successfully demonstrated the correlation between immunointensity of the steroidogenic enzyme and its activity in an evaluation of aromatase in cultured tumor cells. We then showed that immunointensity of P450scc (cholesterol side chain cleavage), an initial step of cortisol biosynthesis, can be also determined per cells by results of cell models, i.e., P450scc antigens attached to the surface of Reacti-Gel HW-65.We studied cell turnover of human adrenal by 3'-OH end labeling method and immunostain of Ki67. We first demonstrated that in human adrenal cortex, cells proliferate in the outer zona fasciculata and die as an apoptosis in the zonal reticularis and glomerulosa, consistent with "cell migration" or considered to play important roles in tumor cell dynamics in adrenocortical neoplasms.We examined expression of Ad4BP (adrenal 4 binding protein) in human adrenal and Ad4BP was demonstrated to be a good biological marker of adrenocortical cell phenotype.