Analysis of growth factors and cytokines in human adrenocortical steroidogenesis

我们从蛋白和mRNA水平检测了转化生长因子-α、表皮生长因子和表皮生长因子受体在人肾上腺皮质及其疾病中的表达。在正常肾上腺和肿瘤肾上腺组织中均可检测到转化生长因子-α的蛋白和mRNA表达，但未检测到转化生长因子受体的表达。然后我们发现，作为皮质醇生物合成的起始步骤，P450scc(胆固醇侧链断裂)的免疫强度也可以通过细胞模型的结果来确定，即P450scc抗原附着在Reacti-Gel HW-65的表面。我们用3‘-OH末端标记法和Ki67免疫染色研究了人肾上腺的细胞周转。我们首次证实在人肾上腺皮质中，细胞在外束状带中增殖，在网状带和肾小球中以细胞凋亡的形式死亡，这与“细胞迁移”一致或被认为在肾上腺皮质肿瘤的细胞动力学中起着重要作用。

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.