Anti-Anaplastic Thyroid Cancer (ATC) Effects and Mechanisms of PLX3397 (Pexidartinib), a Multi-Targeted Tyrosine Kinase Inhibitor (TKI).
Anti-Anaplastic Thyroid Cancer (ATC) Effects and Mechanisms of PLX3397 (Pexidartinib), a Multi-Targeted Tyrosine Kinase Inhibitor (TKI).
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甲状腺癌是甲状腺癌中最常见的恶性肿瘤之一。令人遗憾的是，ATC患者对多种治疗策略的反应很差。因此，必须加强对这种恶性癌症的治疗方法。在本研究中，我们发现Pexidartinib诱导ATC细胞中的ER应激和ROS升高。Pexidartinib给药后ATC中的凋亡细胞和ER应激可分别被ER应激抑制剂和ROS清除剂逆转。此外，Pexidartinib处理诱导Nrf 2（核因子红细胞2相关因子2）在细胞核中蓄积，并降低Nrf 2与Keap-1（Kelch样ECH相关蛋白1）的相互作用，而Nrf 2的敲低增强了Pexidartinib的体外抗ATC作用。此外，Pexidartinib在体内可显著抑制ATC异种移植物生长和增殖，与ML 385（一种Nrf 2抑制剂）联合使用可有效增强Pexidartinib在体内的抗ATC作用。我们的研究结果表明，Pexidartinib是治疗ATC的潜在药物。背景甲状腺未分化癌（Anaplastic thyroid cancer，ATC）是甲状腺恶性肿瘤中最严重的一种，发病率低，缺乏有效的治疗策略和规范的治疗方案。PLX 3397（Pexidartinib）是FDA批准的多靶点酪氨酸激酶抑制剂。本研究旨在探索Pexidartinib对ATC可能的抗增殖活性及其相关分子机制。方法采用CCK-8、LDH释放、集落形成和EdU检测等方法检测细胞活力。流式细胞术（FCM）检测细胞凋亡及细胞周期阻滞的变化。免疫荧光（IF）法检测内质网应激。通过流式细胞术测定活性氧水平。Western印迹分析用于评估与细胞凋亡和ER应激相关的关键分子的变化。建立ATC异种移植模型，并进行免疫组织化学以验证Pexidartinib的体内抗ATC作用。结果Pexidartinib可显著抑制ATC细胞增殖，诱导细胞凋亡和细胞周期阻滞。此外，Pexidartinib可有效诱导ATC细胞中的ER应激和ROS升高，Pexidartinib给药后ATC中的凋亡细胞和ER应激可分别被ER应激抑制剂和ROS清除剂逆转。此外，Pexidartinib处理诱导Nrf 2在细胞核中蓄积，并降低Nrf 2与Keap-1的相互作用，Nrf 2的敲低增强了Pexidartinib的体外抗ATC作用。此外，Pexidartinib在体内可显著抑制ATC异种移植物生长和增殖，与ML 385（一种Nrf 2抑制剂）联合使用可有效增强Pexidartinib在体内的抗ATC作用。结论Pexidartinib是治疗ATC的潜在药物。与Nrf 2抑制剂共同给药是一种有效的协同策略。
Anaplastic thyroid cancer (ATC) is the highest lethal type of thyroid cancer. Regrettably, ATC patients respond poorly to multiple treatment strategies. Therefore, there is imperative to strengthen the therapeutic approach to this vicious form of cancer. In the present study, we found that pexidartinib induces ER stress and elevated ROS in ATC cells. The apoptotic cells, and ER stress in ATC after administration of pexidartinib could be reversed by ER stress inhibitor and ROS scavenger, respectively. Furthermore, pexidartinib treatment induced Nrf2 (Nuclear Factor Erythroid 2–related Factor 2) accumulation in nuclei and reduced the interaction of Nrf2 with Keap-1 (Kelch-like ECH-associated protein 1), while the knockdown of Nrf2 enhanced the anti-ATC effects of pexidartinib in vitro. In addition, pexidartinib significantly inhibits ATC xenografts growth and proliferation in vivo, and the combination of ML385, an Nrf2 inhibitor, potently enhanced the anti-ATC effects of pexidartinib in vivo. Our findings suggest pexidartinib to be a potential agent for treating of ATC. Background Anaplastic thyroid cancer (ATC) is the greatest lethal thyroid neoplasm with a low incidence and lacks an effective treatment strategy and standardized treatment protocol. PLX3397 (Pexidartinib) is an FDA-approved multitarget tyrosine kinase inhibitor. The research is designed to explore the possible anti-proliferative activity of pexidartinib on ATC, as well as its related molecular mechanisms. Methods The cell viability was assessed by CCK-8, LDH release, colony formation, and EdU detection assays. Apoptosis and the alteration on cell cycle arrest were characterized by flow cytometry (FCM). ER stress was evaluated by immunofluorescence (IF). ROS levels were determined by flow cytometry. Western blot assays were conducted to evaluate changes in key molecules related to apoptosis and ER stress. The ATC xenografts model was established, and immunohistochemistry was performed to validate the anti-ATC effects of pexidartinib in vivo. Results Pexidartinib significantly inhibited ATC cell proliferation and induced apoptosis and cell cycle arrest. Moreover, pexidartinib potently induced ER stress and elevated ROS in ATC cells, and the apoptotic cells and ER stress in ATC after administration of pexidartinib could be reversed by an ER stress inhibitor and ROS scavenger, respectively. Furthermore, pexidartinib treatment induced Nrf2 accumulation in nuclei and reduced the interaction of Nrf2 with Keap-1, and knockdown of Nrf2 enhanced the anti-ATC effects of pexidartinib in vitro. In addition, pexidartinib significantly inhibited ATC xenograft growth and proliferation in vivo, and the combination of ML385, an Nrf2 inhibitor, potently enhanced the anti-ATC effects of pexidartinib in vivo. Conclusion Our findings suggest pexidartinib is a potential agent for treating ATC. Co-administration with an Nrf2 inhibitor is an effective synergistic strategy.
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