Anaplastic thyroid carcinoma (ATC) is the most aggressive subtype of thyroid cancer. In this study, we used a three-dimensional in vitro system to evaluate the effect of a dual MEK/Aurora kinase inhibitor, BI-847325 anticancer drug, on several cellular and molecular processes involved in cancer progression. Human ATC cell lines, C643 and SW1736, were grown in alginate hydrogel and treated with IC50 values of BI-847325. The effect of BI-847325 on inhibition of kinases function of MEK1/2 and Aurora kinase B (AURKB) was evaluated via Western blot analysis of phospho-ERK1/2 and phospho-Histone H3 levels. Sodium/iodide symporter (NIS) and thyroglobulin (Tg), as two thyroid-specific differentiation markers, were measured by qRT-PCR as well as flow cytometry and immunoradiometric assay. Apoptosis was assessed by Annexin V/PI flow cytometry and BIM, NFκB1, and NFκB2 expressions. Cell cycle distribution and proliferation were determined via P16, AURKA, and AURKB expressions as well as PI and CFSE flow cytometry assays. Multidrug resistance was evaluated by examining the expression of MDR1 and MRP1. Angiogenesis and invasion were investigated by VEGF expression and F-actin labeling with Alexa Fluor 549 Phalloidin. Western blot results showed that BI-847325 inhibits MEK1/2 and AURKB functions by decreasing phospho-ERK1/2 and phospho-Histone H3 levels. BI-847325 induced thyroid differentiation markers and apoptosis in ATC cell lines. Inversely, BI-847325 intervention decreased multidrug resistance, cell cycle progression, proliferation, angiogenesis, and invasion at the molecular and/or cellular levels. The results of the present study suggest that BI-857,325 might be an effective multi-targeted anticancer drug for ATC treatment.

中文翻译：

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BI-847325，一种选择性双重 MEK 和 Aurora 激酶抑制剂，可减少体外三维培养物中未分化甲状腺癌的侵袭行为

甲状腺未分化癌 (ATC) 是最具侵袭性的甲状腺癌亚型。在这项研究中，我们使用三维体外系统来评估双重 MEK/Aurora 激酶抑制剂 BI-847325 抗癌药物对癌症进展中涉及的几个细胞和分子过程的影响。人类 ATC 细胞系 C643 和 SW1736 在藻酸盐水凝胶中生长，并用 IC50 值的 BI-847325 处理。通过磷酸化 ERK1/2 和磷酸化组蛋白 H3 水平的蛋白质印迹分析评估 BI-847325 对抑制 MEK1/2 和极光激酶 B (AURKB) 激酶功能的影响。钠/碘化物同向转运蛋白 (NIS) 和甲状腺球蛋白 (Tg) 作为两种甲状腺特异性分化标志物，通过 qRT-PCR 以及流式细胞术和免疫放射测定法进行测量。通过 Annexin V/PI 流式细胞术和 BIM、NFκB1、和 NFκB2 的表达。通过 P16、AURKA 和 AURKB 表达以及 PI 和 CFSE 流式细胞术测定确定细胞周期分布和增殖。通过检测 MDR1 和 MRP1 的表达来评估多药耐药性。通过 VEGF 表达和用 Alexa Fluor 549 Phalloidin 标记的 F-肌动蛋白来研究血管生成和侵袭。蛋白质印迹结果显示 BI-847325 通过降低磷酸化 ERK1/2 和磷酸化组蛋白 H3 水平来抑制 MEK1/2 和 AURKB 功能。BI-847325 在 ATC 细胞系中诱导甲状腺分化标志物和细胞凋亡。相反，BI-847325 干预在分子和/或细胞水平上降低了多药耐药性、细胞周期进程、增殖、血管生成和侵袭。本研究的结果表明，BI-857，