Moosavi Fatemeh, Firoozi Roya, Tavakkoli Marjan, Nazari Somayeh, Alipour Alireza, Firuzi Omidreza
Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Medicinal and Natural Products Chemistry Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Toxicology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran.
Biochimie. 2025 Apr;231:73-83. doi: 10.1016/j.biochi.2024.12.006. Epub 2024 Dec 14.
Pancreatic ductal adenocarcinoma (PDAC) remains as one of the most lethal malignancies. c-MET is an important oncogenic kinase involved in PDAC progression. We determined the anticancer effect of c-MET inhibitors, crizotinib and cabozantinib, combined with chemotherapeutic agents, doxorubicin, oxaliplatin and gemcitabine, against different PDAC and a lung adenocarcinoma cell line expressing different levels of c-MET. MTT assay was performed to assess cell growth inhibition. Synergistic combinations were evaluated in spheroid cultures, while apoptosis was determined through Hoechst33258 staining. The effect of drug combinations on cell cycle and apoptosis induction was examined by RNase/PI flow cytometric assay. We also evaluated reactive oxygen species (ROS) levels using 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay to explore the possible mechanisms contributing to synergism. Combination of crizotinib or cabozantinib with doxorubicin exhibited synergistic effects in c-MET overexpressing cells. Conversely, combinations of c-MET inhibitors with other agents were additive or even antagonistic. Combination index (CI) values calculated with Calcusyn software were 0.631-0.730 for crizotinib and 0.542-0.746 for cabozantinib co-administered with doxorubicin. These synergistic combinations showed significant spheroid growth inhibition and apoptosis induction in Suit-2, c-MET dependent PDAC cells. These combinations also significantly increased the number of cells in both apoptotic sub-G1 phase and the G2/M phase compared to single-drug treatment. Increased ROS production seemed to be a possible mechanism underlying synergism. In conclusion, c-MET inhibitors synergize with DNA damaging agent, doxorubicin, in cancer cells with c-MET overexpression, indicating that these combination therapies may be a promising cancer therapeutic strategy.
胰腺导管腺癌（PDAC）仍然是最致命的恶性肿瘤之一。c-MET是一种参与PDAC进展的重要致癌激酶。我们确定了c-MET抑制剂克唑替尼和卡博替尼与化疗药物阿霉素、奥沙利铂和吉西他滨联合使用对不同的PDAC细胞系和表达不同水平c-MET的肺腺癌细胞系的抗癌效果。进行MTT试验以评估细胞生长抑制情况。在球体培养中评估协同组合，同时通过Hoechst33258染色确定细胞凋亡情况。通过核糖核酸酶/碘化丙啶流式细胞术检测药物组合对细胞周期和凋亡诱导的影响。我们还使用2'，7'-二氯荧光素二乙酸酯（DCFH-DA）试验评估活性氧（ROS）水平，以探索导致协同作用的可能机制。克唑替尼或卡博替尼与阿霉素联合使用在c-MET过表达细胞中表现出协同作用。相反，c-MET抑制剂与其他药物的组合是相加的甚至是拮抗的。用Calcusyn软件计算的联合指数（CI）值，克唑替尼与阿霉素联合使用时为0.631 - 0.730，卡博替尼与阿霉素联合使用时为0.542 - 0.746。这些协同组合在Suit-2（c-MET依赖性PDAC细胞）中显示出显著的球体生长抑制和凋亡诱导作用。与单药治疗相比，这些组合还显著增加了凋亡亚G1期和G2/M期的细胞数量。ROS产生增加似乎是协同作用的一个可能机制。总之，在c-MET过表达的癌细胞中，c-MET抑制剂与DNA损伤剂阿霉素协同作用，表明这些联合疗法可能是一种有前景的癌症治疗策略。
