The effects of the standardized extracts of Ginkgo biloba on steroidogenesis pathways and aromatase activity in H295R human adrenocortical carcinoma cells.

Q3 Medicine Environmental Health and Toxicology Pub Date : 2016-05-09 eCollection Date: 2016-01-01 DOI:10.5620/eht.e2016010

Mijie Kim, Yong Joo Park, Huiyeon Ahn, Byeonghak Moon, Kyu Hyuck Chung, Seung Min Oh

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引用次数: 17

Abstract

Objectives: Aromatase inhibitors that block estrogen synthesis are a proven first-line hormonal therapy for postmenopausal breast cancer. Although it is known that standardized extract of Ginkgo biloba (EGb761) induces anti-carcinogenic effects like the aromatase inhibitors, the effects of EGb761 on steroidogenesis have not been studied yet. Therefore, the effects of EGb761 on steroidogenesis and aromatase activity was studied using a H295R cell model, which was a good in vitro model to predict effects on human adrenal steroidogenesis.

Methods: Cortisol, aldosterone, testosterone, and 17β-estradiol were evaluated in the H295R cells by competitive enzyme-linked immunospecific assay after exposure to EGb761. Real-time polymerase chain reaction were performed to evaluate effects on critical genes in steroid hormone production, specifically cytochrome P450 (CYP11/ 17/19/21) and the hydroxysteroid dehydrogenases (3β-HSD2 and 17β-HSD1/4). Finally, aromatase activities were measured with a tritiated water-release assay and by western blotting analysis.

Results: H295R cells exposed to EGb761 (10 and 100 μg/mL) showed a significant decrease in 17β-estradiol and testosterone, but no change in aldosterone or cortisol. Genes (CYP19 and 17β-HSD1) related to the estrogen steroidogenesis were significantly decreased by EGb761. EGb761 treatment of H295R cells resulted in a significant decrease of aromatase activity as measured by the direct and indirect assays. The coding sequence/ Exon PII of CYP19 gene transcript and protein level of CYP19 were significantly decreased by EGb761.

Conclusions: These results suggest that EGb761 could regulate steroidogenesis-related genes such as CYP19 and 17β-HSD1, and lead to a decrease in 17β-estradiol and testosterone. The present study provides good information on potential therapeutic effects of EGb761 on estrogen dependent breast cancer.

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银杏标准化提取物对H295R人肾上腺皮质癌细胞甾体生成途径及芳香化酶活性的影响。

目的:阻断雌激素合成的芳香酶抑制剂是绝经后乳腺癌的一线激素治疗方法。虽然已知银杏标准化提取物(EGb761)具有与芳香化酶抑制剂一样的抗癌作用，但EGb761对甾体生成的影响尚未得到研究。因此，我们使用H295R细胞模型来研究EGb761对类固醇生成和芳香酶活性的影响，H295R细胞模型是一个很好的体外模型，可以预测对人肾上腺类固醇生成的影响。方法:应用竞争性酶联免疫特异法测定暴露于EGb761后H295R细胞中皮质醇、醛固酮、睾酮和17β-雌二醇的水平。采用实时聚合酶链反应(Real-time polymerase chain reaction)评估对类固醇激素生成关键基因的影响，特别是细胞色素P450 (CYP11/ 17/19/21)和羟基类固醇脱氢酶(3β-HSD2和17β-HSD1/4)。最后，用氚化水释放法和western blotting法测定芳香酶活性。结果:EGb761(10和100 μg/mL)作用于H295R细胞后，17β-雌二醇和睾酮水平显著降低，醛固酮和皮质醇水平无明显变化。EGb761显著降低了与雌激素甾体生成相关的基因(CYP19和17β-HSD1)。通过直接和间接测定，EGb761处理H295R细胞导致芳香酶活性显著降低。EGb761显著降低了CYP19基因转录物的编码序列/外显子PII和CYP19蛋白水平。结论:上述结果提示EGb761可调控激素生成相关基因CYP19、17β-HSD1，导致17β-雌二醇和睾酮水平降低。本研究为EGb761对雌激素依赖性乳腺癌的潜在治疗作用提供了良好的信息。

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