Inhibition of Src tyrosine kinase stimulates adrenal androgen production
Inhibition of Src tyrosine kinase stimulates adrenal androgen production
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DOI：
10.1677/jme.0.0300287
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发表时间：
2003-06-01
影响因子：
3.5
通讯作者：
Rainey, WE
中科院分区：
文献类型：
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作者：
Sirianni, R;Carr, BR;Rainey, WE
灵长类动物肾上腺的一个独特特征是能够产生 19 碳类固醇，通常称为肾上腺雄激素。尽管很明显，主要的人类肾上腺雄激素脱氢表雄酮（DHEA）和硫酸脱氢表雄酮（DHEA-S）几乎仅在肾上腺网状层中产生，但人们对调节产生的机制知之甚少。在此，我们测试了酪氨酸激酶 Src 家族参与肾上腺雄激素产生调节的假设。使用NC1-H295R人肾上腺细胞系和培养的原代人肾上腺细胞来研究肾上腺雄激素的产生和参与类固醇生成的酶的表达。为了检查 Src 酪氨酸激酶的作用，用 PP2（一种特定的 Src 抑制剂）处理细胞。或者，用含有显性失活形式的 Src 的表达载体转染肾上腺细胞。在两种肾上腺细胞模型中，PP2 治疗抑制基础皮质醇的产生，同时显着增加 DHEA 和 DHEA-S（统称为 DHEA(S)）的产生。 Western 分析显示，PP2 对类固醇生成的影响伴随着类固醇生成急性调节 (StAR) 蛋白合成的快速诱导而发生。 PP2 治疗还增加了 StAR、胆固醇侧链裂解 (CYP11A) 和 17α-羟化酶/17,20-裂解酶 (CYP17) 的 mRNA 水平。用 cAMP 激动剂环单磷酸二丁酰腺苷 (dbcAMP) 处理肾上腺细胞，刺激皮质醇和 DHEA(S) 的产生。然而，用 PP2 和 dbcAMP 组合处理肾上腺细胞可增强 DHEA(S) 的产生，同时抑制皮质醇的产生。在 dbcAMP 治疗期间，PP2 能够增强 CYP17 的表达并抑制 2 型 3β-羟基类固醇脱氢酶 (HSD3B2) 水平的诱导。增加 CYP17 与 HSD3B2 的比例可能会促进使用类固醇前体来生产 DHEA(S)，而不是皮质醇。综合这些数据表明，Src 酪氨酸激酶的抑制导致肾上腺细胞通过产生 DHEA(S) 和表达类固醇生成酶而采用网状表型。
A unique characteristic of the primate adrenal is the ability to produce 19-carbon steroids, often called the adrenal androgens. Although it is clear that the major human adrenal androgens, dehydroepiandrosterone (DHEA) and DHEA sulfate (DHEA-S), are produced almost solely in the adrenal reticularis, the mechanisms regulating production are poorly understood. Herein, we tested the hypothesis that the Src family of tyrosine kinases are involved in the regulation of adrenal androgen production. The NCl-H295R human adrenal cell line and primary human adrenal cells in culture were used to study adrenal androgen production and expression of enzymes involved in steroidogenesis. To examine the role of Src tyrosine kinase, cells were treated with PP2, a specific Src inhibitor. Alternatively, adrenal cells were transfected with an expression vector containing a dominant-negative form of Src. PP2 treatment inhibited basal cortisol production while significantly increasing the production of DHEA and DHEA-S (together referred to as DHEA(S)) in both adrenal cell models. The effect of PP2 on steroidogenesis occurred along with a rapid induction of steroidogenic acute regulatory (StAR) protein synthesis as revealed by Western analysis. Treatment with PP2 also increased mRNA levels for StAR, and cholesterol side-chain cleavage (CYP11A) and 17alpha-hydroxylase/17,20-lyase (CYP17) enzymes. Treatment of adrenal cells with the cAMP agonist dibutyryladenosine cyclic monophosphate (dbcAMP), stimulated the production of cortisol and DHEA(S). However, treatment of adrenal cells with a combination of PP2 and dbcAMP enhanced the production of DHEA(S) while inhibiting cortisol production. During dbcAMP treatment PP2 was able to augment the expression of CYP17 and to inhibit the induction of 3beta-hydroxysteroid dehydrogenase type 2 (HSD3B2) levels. Increasing the CYP17 to HSD3B2 ratio is likely to promote the use of steroid precursors for the production of DHEA(S) and not for cortisol. Taken together these data suggest that the inhibition of Src tyrosine kinases causes adrenal cells to adopt a reticularis phenotype both by the production of DHEA(S) and by the steroidogenic enzymes expressed.