Laboratory of Pharmacology, Chulabhorn Research Institute, Bangkok 10210, Thailand.
Int J Oncol. 2018 Jul;53(1):177-188. doi: 10.3892/ijo.2018.4375. Epub 2018 Apr 24.
Cholangiocarcinoma (CCA) is a malignant tumor of the biliary epithelium associated with Opisthorchis viverrini, primary sclerosing cholangitis and hepatitis viral infection. In the global population, men have higher incidence rates for CCA than women; thus, a gender disparity in the progression of chronic inflammation of the biliary duct leading to malignancy may involve the effects of estrogen (E2). Genistein (GE), a prominent phytoestrogen found in soy products, is an estrogen receptor β (ERβ) agonist and a tyrosine kinase inhibitor. The present study investigated the effects of GE on the growth of CCA cells by cell viability assay. The effects on signaling proteins were detected by western blot analysis and ELISA. Gene expression was examined by RT-qPCR. Two human intrahepatic CCA cell lines, HuCCA‑1 and RMCCA‑1, were utilized. GE (50‑200 µM) reduced the viability of the two cell lines, and also inhibited the activation of epidermal growth factor receptor (EGFR) and AKT, as evidenced by decreasing protein levels of phosphorylated (p)-EGFR (Tyr1173) and p‑AKT (Ser473), respectively. GE altered the mitogen‑activated protein kinase signaling cascade by mediating decreased protein levels of p‑p38 and increased protein levels of p‑ERK1/2. GE significantly decreased the levels of interleukin 6 (IL6) and induced the expression of inducible nitric oxide synthase (iNOS). GE also downregulated the expression of p‑ERα (Ser118) protein and ERα mRNA levels. Finally, GE induced the downregulation of the protein levels of ERβ. Of note, E2 deprivation potentiated the GE-induced reduction of p‑EGFR (Tyr1173) and total AKT proteins and production of IL6, and mediated the downregulation of GE-induced iNOS protein. In conclusion, GE inhibited the growth of human CCA cell lines by reducing the activation of EGFR and AKT, and by attenuating the production of IL6. E2 and ER were also involved in the growth-inhibitory effect of GE in CCA cells.
胆管癌（CCA）是一种与华支睾吸虫、原发性硬化性胆管炎和肝炎病毒感染相关的胆道上皮恶性肿瘤。在全球人群中，男性 CCA 的发病率高于女性；因此，慢性胆管炎症向恶性肿瘤进展的性别差异可能涉及雌激素（E2）的影响。染料木黄酮（GE）是一种存在于大豆制品中的主要植物雌激素，是一种雌激素受体β（ERβ）激动剂和酪氨酸激酶抑制剂。本研究通过细胞活力测定法研究了 GE 对 CCA 细胞生长的影响。通过 Western blot 分析和 ELISA 检测信号蛋白的作用。通过 RT-qPCR 检查基因表达。使用两种人肝内 CCA 细胞系 HuCCA-1 和 RMCCA-1。GE（50-200μM）降低了两种细胞系的活力，并通过分别降低磷酸化（p）-表皮生长因子受体（EGFR）（Tyr1173）和 p-AKT（Ser473）的蛋白水平来抑制 EGFR 和 AKT 的激活。GE 通过调节 p-p38 和增加 p-ERK1/2 的蛋白水平来改变丝裂原激活的蛋白激酶信号级联。GE 显著降低白细胞介素 6（IL6）的水平并诱导诱导型一氧化氮合酶（iNOS）的表达。GE 还下调了 p-ERα（Ser118）蛋白和 ERα mRNA 水平的表达。最后，GE 诱导 ERβ 蛋白水平下调。值得注意的是，E2 剥夺增强了 GE 诱导的 p-EGFR（Tyr1173）和总 AKT 蛋白减少以及 IL6 的产生，并介导了 GE 诱导的 iNOS 蛋白下调。总之，GE 通过降低 EGFR 和 AKT 的激活以及减弱 IL6 的产生来抑制人 CCA 细胞系的生长。E2 和 ER 也参与了 GE 在 CCA 细胞中的生长抑制作用。
