Departments of Otolaryngology, Head & Neck Surgery, LSU Health Shreveport, 1501 Kings Highway, Shreveport, LA 71103, USA.
Departments of Molecular and Cellular Physiology, LSU Health Shreveport, 1501 Kings Highway, Shreveport, LA 71103, USA.
Int J Mol Sci. 2021 Feb 16;22(4):1961. doi: 10.3390/ijms22041961.
Anaplastic thyroid cancer (ATC) is one of the most lethal malignancies with a median survival time of about 4 months. Currently, there is no effective treatment, and the development of new therapies is an important and urgent issue for ATC patients. YM155 is a small molecule that was identified as the top candidate in a high-throughput screen of small molecule inhibitors performed against a panel of ATC cell lines by the National Cancer Institute. However, there were no follow-up studies investigating YM155 in ATC. Here, we determined the effects of YM155 on ATC and human primary benign thyroid cell (PBTC) survival with alamarBlue assay. Our data show that YM155 inhibited proliferation of ATC cell lines while sparing normal thyroid cells, suggesting a high therapeutic window. YM155-induced DNA damage was detected by measuring phosphorylation of γ-H2AX as a marker for DNA double-strand breaks. The formamidopyrimidine-DNA glycosylase (FPG)-modified alkaline comet assay in conjunction with reactive oxygen species (ROS) assay and glutathione (GSH)/glutathione (GSSG) assay suggests that YM155-mediated oxidative stress contributes to DNA damage. In addition, we provide evidence that YM155 causes cell cycle arrest in S phase and in the G2/M transition and causes apoptosis, as seen with flow cytometry. In this study, we show for the first time the multiple effects of YM155 in ATC cells, furthering a potential therapeutic approach for ATC.
间变性甲状腺癌 (ATC) 是最致命的恶性肿瘤之一,中位生存时间约为 4 个月。目前尚无有效的治疗方法,开发新的治疗方法是 ATC 患者的重要而紧迫的问题。YM155 是一种小分子,在国立癌症研究所对一系列 ATC 细胞系进行的小分子抑制剂高通量筛选中被确定为首选候选药物。然而,没有后续研究调查 YM155 在 ATC 中的作用。在这里,我们通过 alamarBlue 测定法确定了 YM155 对 ATC 和人原代良性甲状腺细胞 (PBTC) 存活的影响。我们的数据表明,YM155 抑制 ATC 细胞系的增殖,同时保留正常甲状腺细胞,表明治疗窗口较高。通过测量 γ-H2AX 的磷酸化作为 DNA 双链断裂的标志物来检测 YM155 诱导的 DNA 损伤。与活性氧 (ROS) 测定和谷胱甘肽 (GSH)/谷胱甘肽 (GSSG) 测定相结合的 formamidopyrimidine-DNA 糖基化酶 (FPG) 修饰的碱性彗星试验表明,YM155 介导的氧化应激导致 DNA 损伤。此外,我们提供的证据表明,YM155 导致细胞周期停滞在 S 期和 G2/M 过渡,并通过流式细胞术观察到细胞凋亡。在这项研究中,我们首次展示了 YM155 在 ATC 细胞中的多种作用,为 ATC 的潜在治疗方法提供了进一步的证据。