Cellular and Molecular Endocrine Research Center, Research Institute for Endocrine Sciences, Shahid Beheshti University of Medical Sciences, No.23, Yemen St, Aarabi Street, 193954763, Tehran, Iran.
Department of Biochemistry, Falavarjan Branch Islamic Azad University, Isfahan, Iran.
J Endocrinol Invest. 2024 Nov;47(11):2809-2821. doi: 10.1007/s40618-024-02377-4. Epub 2024 May 2.
The alternative manner of iodide and glucose uptake found in different types of thyroid cancer, referred to flip-flop. ATC cells indicate low iodide uptake and high glucose uptake, which lack the morphology and genetic characteristics of well-differentiated tumors and become increasingly invasive. Importance placed on the discovery of innovative multi-targeted medicines to suppress the dysregulated signaling in cancer. In this research, we aimed to clarify molecular mechanism of Rutin as a phytomedicine on anaplastic thyroid cancer cell line based on iodide and glucose uptake.
The MTT test was employed to test cell viability. Iodide uptake assay was performed using a spectrophotometric assay to determine iodide uptake in SW1736 cells based on Sandell-Kolthoff reaction. For glucose uptake detection, ''GOD-PAP'' enzymatic colorimetric assay was applied to measure the direct glucose levels inside of the cells. Determination of NIS, GLUT1 and 3 mRNA expression in SW1736 cells was performed by qRT-PCR. Determination of NIS, GLUT1 and 3 protein levels in SW1736 cells was performed by western blotting.
According to our results, Rutin inhibited the viability of SW1736 cells in a time- and dose-dependent manner. Quantitative Real-time RT-PCR analysis exposed that NIS mRNA levels were increased in Rutin treated group compared to the control group. Accordingly, western blot showed high expression of NIS protein and low expression of GLUT 1 and 3 in Rutin treated SW1736 cell line. Rutin increased iodide uptake and decreased glucose uptake in thyroid cancer cell line SW1736 compared to control group.
Multiple mechanisms point to Rutin's role as a major stimulator of iodide uptake and inhibitor of glucose uptake, including effects at the mRNA and protein levels for both NIS and GLUTs, respectively. Here in, we described the flip-flop phenomenon as a possible therapeutic target for ATC. Moreover, Rutin is first documented here as a NIS expression inducer capable of restoring cell differentiation in SW1736 cell line. It also be concluded that GLUTs as metabolic targets can be blocked specifically by Rutin for thyroid cancer prevention and treatment.
在不同类型的甲状腺癌中发现了碘和葡萄糖摄取的替代方式，称为翻转。ATC 细胞表现出低碘摄取和高葡萄糖摄取，缺乏分化良好肿瘤的形态和遗传特征，并且变得越来越具有侵袭性。人们越来越重视发现创新的多靶向药物来抑制癌症中失调的信号转导。在这项研究中，我们旨在基于碘和葡萄糖摄取阐明芦丁作为植物药对间变性甲状腺癌细胞系的作用机制。
采用 MTT 试验检测细胞活力。通过分光光度法测定 SW1736 细胞的碘摄取，根据桑德尔-科尔托夫反应（Sandell-Kolthoff reaction）测定碘摄取。为了检测葡萄糖摄取，应用“GOD-PAP”酶比色法测量细胞内的直接葡萄糖水平。通过 qRT-PCR 测定 SW1736 细胞中 NIS、GLUT1 和 3 mRNA 的表达。通过 Western blot 测定 SW1736 细胞中 NIS、GLUT1 和 3 蛋白水平。
根据我们的结果，芦丁以时间和剂量依赖的方式抑制 SW1736 细胞的活力。定量实时 RT-PCR 分析表明，芦丁处理组的 NIS mRNA 水平高于对照组。相应地，Western blot 显示芦丁处理的 SW1736 细胞系中 NIS 蛋白高表达，GLUT1 和 3 低表达。芦丁增加甲状腺癌细胞系 SW1736 中的碘摄取并减少葡萄糖摄取，与对照组相比。
多种机制表明芦丁作为碘摄取的主要刺激剂和葡萄糖摄取的抑制剂的作用，包括分别在 NIS 和 GLUTs 的 mRNA 和蛋白水平上的作用。在这里，我们将翻转现象描述为 ATC 的可能治疗靶点。此外，芦丁首次被证明是一种能够在 SW1736 细胞系中恢复细胞分化的 NIS 表达诱导剂。还可以得出结论，GLUTs 作为代谢靶点可以被芦丁特异性阻断，用于预防和治疗甲状腺癌。
