Wang Ruikai, Li Amin, Liu Jiachang, Fang Ming, Zhu Yan, Huang Juan, Liu Yinjie, Huo Long, You Qinghai
Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Anhui Medical University, Hefei, China.
Pulmonary and Critical Care Medicine, First Affiliated Hospital, Anhui University of Science & Technology (Huainan First People's Hospital), Huainan, China.
J Chemother. 2023 Apr;35(2):95-103. doi: 10.1080/1120009X.2022.2045826. Epub 2022 Mar 3.
Cisplatin, as a first-line chemotherapy drug for advanced wild-type epidermal growth factor receptor (wtEGFR) non-small cell lung cancer (NSCLC), often loses effectiveness because of acquired drug resistance. We found that ataxia-telangiectasia mutated (ATM), ataxia-telangiectasia and Rad3-related (ATR) and DNA-dependent protein kinase catalytic subunit (DNA-PKcs) of DNA repair kinases and signal transduction molecules, protein kinase B (AKT)/target mammalian target of rapamycin (mTOR), were significantly phosphorylated in cisplatin-resistant wtEGFR NSCLC cell lines (H358R and A549R) than in their parental cells. Also, BEZ235 (dual phosphatidylinositol-3-kinase (PI3K)/mTOR inhibitor) significantly decreased the phosphorylation levels of these kinases/proteins, as detected by Western blot analysis. In H358R and A549R cells, the results of indirect immunofluorescence, single-cell gel electrophoresis, flow cytometry, methylthiazolyldiphenyl-tetrazolium bromide, clone formation assay, and scratch healing experiment showed that BEZ235 enhanced cisplatin-induced DNA damage and cell apoptosis, and effectively inhibited cellular proliferation/migration when combined with cisplatin. The data indicated that the abnormal activation of ATM/ATR/DNA-PKcs kinases and AKT/mTOR pathway might induce wtEGFR NSCLC cell resistance to cisplatin. The effects of the combination of BEZ235 and cisplatin suggested that BEZ235 should be considered as a combination therapy for patients with cisplatin-resistant wtEGFR NSCLC.
顺铂作为晚期野生型表皮生长因子受体（wtEGFR）非小细胞肺癌（NSCLC）的一线化疗药物，常因获得性耐药而失效。我们发现，在顺铂耐药的wtEGFR NSCLC细胞系（H358R和A549R）中，DNA修复激酶和信号转导分子的共济失调毛细血管扩张突变（ATM）、共济失调毛细血管扩张和Rad3相关（ATR）以及DNA依赖性蛋白激酶催化亚基（DNA-PKcs）、蛋白激酶B（AKT）/雷帕霉素靶蛋白（mTOR）的磷酸化水平显著高于其亲本细胞。此外，通过蛋白质印迹分析检测发现，BEZ235（双磷脂酰肌醇-3-激酶（PI3K）/mTOR抑制剂）可显著降低这些激酶/蛋白的磷酸化水平。在H358R和A549R细胞中，间接免疫荧光、单细胞凝胶电泳、流式细胞术、甲基噻唑基二苯基四氮唑溴盐、克隆形成试验和划痕愈合实验结果表明，BEZ235增强了顺铂诱导的DNA损伤和细胞凋亡，并在与顺铂联合使用时有效抑制了细胞增殖/迁移。数据表明ATM/ATR/DNA-PKcs激酶和AKT/mTOR通路的异常激活可能诱导wtEGFR NSCLC细胞对顺铂产生耐药性。BEZ235与顺铂联合使用的效果表明，BEZ235应被视为顺铂耐药wtEGFR NSCLC患者的联合治疗药物。
