Dose-dependence in acquisition of drug tolerant phenotype and high RYK expression as a mechanism of osimertinib tolerance in lung cancer
Dose-dependence in acquisition of drug tolerant phenotype and high RYK expression as a mechanism of osimertinib tolerance in lung cancer
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DOI：
10.1016/j.lungcan.2021.02.017
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发表时间：
2021-02-22
期刊：
影响因子：
5.3
通讯作者：
Mitsudomi, Tetsuya
中科院分区：
文献类型：
--
作者：
Ohara, Shuta;Suda, Kenichi;Mitsudomi, Tetsuya
目标：在EGFR酪氨酸激酶抑制剂治疗携带EGFR突变的非小细胞肺癌（NSCLC）期间，获得性耐药的出现几乎不可避免。药物耐受是酪氨酸激酶抑制剂治疗早期阶段药物不敏感的可逆状态，被认为是疾病复发的基础。材料与方法：本研究选用5株EGFR突变的NSCLC细胞株，分别用MTT法、MTT法和MTT法检测EGFR突变对NSCLC耐药的影响。我们通过用奥希替尼（600 nM）或阿法替尼（60 nM）处理72 h建立了耐药细胞（DTC）。结果：奥希替尼能诱导PC9、HCC 4006和H1975细胞产生DTCs，阿法替尼能诱导PC9细胞产生DTCs。我们观察到诱导DTC需要高药物浓度，当使用更高剂量的阿法替尼（>180 nM）时，HCC 4006细胞变得耐受。Osimertinib诱导的肝癌4006细胞中受体样酪氨酸激酶（RYK）表达增加，siRNA介导的RYK敲低抑制了Osimertinib诱导的肝癌4006细胞的增殖。结论：Osimertinib诱导的肝癌4006细胞中RYK表达增加是Osimertinib耐药的机制。
Objective: Emergence of acquired resistance is almost inevitable during EGFR-tyrosine kinase inhibitor therapy for non-small-cell lung cancer (NSCLC) harboring EGFR mutations. Drug tolerance, a reversible state of drug insensitivity in the early phases of tyrosine kinase inhibitor therapy, is considered to serve as the basis of recurrent disease. Therefore, it is important to elucidate the molecular mechanisms of drug tolerance.Materials and methods: Five EGFR-mutated NSCLC cell lines were used in this study. We established drug-tolerant cells (DTCs) via 72 h treatment with osimertinib (600 nM) or afatinib (60 nM). Acquisition of drug tolerance was evaluated by growth inhibitory assay, and the molecular mechanisms of drug tolerance were analyzed by phospho-RTK array.Results: DTCs were successfully induced in PC9, HCC4006, and H1975 cells against osimertinib and in PC9 cells against afatinib. We observed that a high drug concentration was required to induce DTCs, and HCC4006 cells become tolerant when a higher dose of afatinib (>180 nM) was used. In the analysis of HCC4006 DTCs against osimertinib, we observed increased receptor-like tyrosine kinase (RYK) expression, and siRNA-mediated RYK knockdown inhibited the proliferation of DTCs.Conclusions: These results suggest that induction of DTCs is dose-dependent, and increased RYK expression was the mechanism of drug tolerance in HCC4006 cells against osimertinib.