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Analysis of growth factors and cytokines in human adrenocortical steroidogenesis

人肾上腺皮质类固醇生成中生长因子和细胞因子的分析

基本信息

批准号：
06670181
负责人：
SASANO Hironobu
金额：
$ 1.22万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

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.

我们检测了tgf - α、EGF和EGFR在人肾上腺皮质及其疾病中的表达，包括蛋白和mRNA水平。在正常和肿瘤肾上腺中均检测到tgf - α蛋白和mRNA水平的表达，但未检测到EGF。EGFR免疫反应性比tgf - α免疫反应性分布更广泛，tgf - α和EGF被认为参与包括肾上腺皮质恶性肿瘤在内的生物学功能。我们在培养的肿瘤细胞中成功地证明了类固醇生成酶的免疫强度与其活性之间的相关性。然后，我们发现P450scc（胆固醇侧链切割）的免疫强度（皮质醇生物合成的第一步）也可以通过细胞模型的结果来确定每个细胞，即附着在reactii - gel HW-65表面的P450scc抗原。采用3'-OH末端标记法和Ki67免疫染色法研究了人肾上腺细胞的转换。我们首先证明，在人肾上腺皮质中，细胞在外束带增殖，在网状带和肾小球中凋亡，与“细胞迁移”一致，或被认为在肾上腺皮质肿瘤的肿瘤细胞动力学中起重要作用。我们检测了Ad4BP（肾上腺4结合蛋白）在人肾上腺中的表达，Ad4BP被证明是肾上腺皮质细胞表型的一个很好的生物学标记。