Ketoconazole Treatment Decreases the Viability of Immortalized Pituitary Cell Lines Associated with an Increased Expression of Apoptosis-Related Genes and Cell Cycle Inhibitors
Ketoconazole Treatment Decreases the Viability of Immortalized Pituitary Cell Lines Associated with an Increased Expression of Apoptosis-Related Genes and Cell Cycle Inhibitors
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DOI：
10.1111/jne.12277
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发表时间：
2015-07-01
影响因子：
3.2
通讯作者：
Bronstein, M. D.
中科院分区：
文献类型：
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作者：
Guzzo, M. F.;Carvalho, L. R.;Bronstein, M. D.
酮康唑最初是作为抗真菌剂开发的，是一种有效的肾上腺类固醇生成抑制剂，因此已用于治疗库欣病。令人惊讶的是，酮康唑治疗期间皮质醇水平的降低并不伴随着预期的血浆促肾上腺皮质激素（ACTH）升高，在失去负皮质醇反馈的促皮质细胞，这表明酮康唑对这些细胞的直接影响。为了表征酮康唑的直接作用，我们使用垂体肿瘤细胞系AtT-20（分泌ACTH）、GH 3（分泌生长激素和催乳素）和T3.1（分泌β亚基）评价了其对细胞活力的体外作用，我们还通过定量逆转录聚合酶链反应（qRT-PCR）测定了参与细胞凋亡和DNA复制的基因的表达水平。我们还评估了酮康唑治疗期间AtT-20细胞中的ACTH水平。我们观察到酮康唑浓度依赖性降低垂体细胞活力和减少ACTH水平的AtT-20细胞后，去除药物。我们还观察到细胞死亡受体（例如Fas，肿瘤坏死因子受体）和半胱天冬酶（例如，半胱天冬酶-6、半胱天冬酶-7、半胱天冬酶-9），表明细胞凋亡途径的激活。此外，我们观察到GH 3细胞中细胞周期抑制剂p21和p27的基因表达增加，T3.1细胞中p21的表达增加。总之，我们的研究结果表明，酮康唑显着降低垂体瘤细胞系中的细胞活力，在浓度依赖性的方式，并与细胞凋亡和细胞周期调控相关的基因表达的增加。
Ketoconazole, which was initially developed as an antifungal agent, is a potent inhibitor of adrenal steroidogenesis and has therefore been used in the management of Cushing's disease. Surprisingly, the reduction of cortisol levels during ketoconazole treatment is not accompanied by the expected elevation in plasma adrenocorticotrophic hormone (ACTH) at the loss of negative cortisol feedback from corticotrophic cells, suggesting a direct effect of ketoconazole on these cells. To characterize the direct effects of ketoconazole, we evaluated its invitro effect on cell viability using the pituitary tumoural cell lines AtT-20 (which secretes ACTH), GH3 (which secretes growth hormone and prolactin) and T3.1 (which secretes -subunit) and we also determined the expression levels of genes involved in apoptosis and DNA replication by the quantitative reverse transcription polymerase chain reaction (qRT-PCR). We also evaluated ACTH levels in AtT-20 cells during ketoconazole treatment. We observed a ketoconazole concentration-dependent decrease in pituitary cell viability and reduced ACTH levels in AtT-20 cells after removal of the drug. We also observed increased expression of cell death receptors (e.g. Fas, tumour necrosis factor receptor) and caspases (e.g., caspase-6, caspase-7, caspase-9), suggesting activation of the apoptosis pathway. In addition, we observed increased gene expression of the cell cycle inhibitors p21 and p27 in GH3 cells and increased expression of p21 in T3.1 cells. In conclusion, our findings suggest that ketoconazole significantly reduces cell viability in a concentration-dependent manner in pituitary tumour cell lines and is associated with an increase in apoptosis- and cell cycle regulation-related gene expression.