奥希替尼在 EGFR-T790M 非小细胞肺癌耐药细胞中诱导增殖和迁移的相反作用。
Osimertinib Induces the Opposite Effect of Proliferation and Migration in the Drug Resistance of EGFR-T790M Non-small Cell Lung Cancer Cells.
机构信息
Institute of Molecular Medicine and Bioengineering, National Yang Ming Chiao Tung University, Hsinchu, Taiwan.
Department of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsinchu, Taiwan.
出版信息
Anticancer Agents Med Chem. 2023;23(11):1309-1319. doi: 10.2174/1871520623666230223111217.
BACKGROUND
Lung cancer has become one of the leading causes of cancer incidence and mortality worldwide. Non-small cell lung carcinoma (NSCLC) is the most common type among all lung cancer cases. NSCLC patients contained high levels of activating epidermal growth factor receptor (EGFR) mutations, such as exon 19 deletion, L858R and T790M. Osimertinib, a third-generation of EGFR tyrosine kinase inhibitor (EGFR-TKI), has therapeutic efficacy on the EGFR-T790M mutation of NSCLC patients; however, treatment of osimertinib still can induce drug resistance in lung cancer patients. Therefore, investigation of the drug resistance mechanisms of osimertinib will provide novel strategies for lung cancer therapy.
METHODS
The H1975OR osimertinib-resistant cell line was established by prolonged exposure with osimertinib derived from the H1975 cells. The cell proliferation ability was evaluated by the cell viability and cell growth assays. The cell migration ability was determined by the Boyden chamber assays. The differential gene expression profile was analyzed by genome-wide RNA sequencing. The protein expression and location were analyzed by western blot and confocal microscopy.
RESULTS
In this study, we established the osimertinib-resistant H1975 (T790M/L858R) cancer cells, named the H1975OR cell line. The cell growth ability was decreased in the H1975OR cells by comparison with the H1975 parental cells. Conversely, the cell migration ability was elevated in the H1975OR cells. We found the differential gene expression profile of cell proliferation and migration pathways between the H1975OR and H1975 parental cells. Interestingly, the protein levels of phospho-EGFR, PD-L1, E-cadherin and β-catenin were decreased, but the survivin and N-cadherin proteins were increased in the H1975OR drug-resistant cells.
CONCLUSION
Osimertinib induces the opposite effect of proliferation and migration in the drug resistance of EGFRT790M lung cancer cells. We suggest that differential gene and protein expressions in the cell proliferation and migration pathways may mediate the drug resistance of osimertinib in lung cancer cells. Understanding the molecular drugresistant mechanisms of proliferation and migration pathways of osimertinib may provide novel targets and strategies for the clinical treatment of EGFR-TKIs in lung cancer patients.
背景
肺癌已成为全球癌症发病率和死亡率的主要原因之一。非小细胞肺癌（NSCLC）是所有肺癌病例中最常见的类型。非小细胞肺癌患者存在高水平的激活表皮生长因子受体（EGFR）突变，如外显子 19 缺失、L858R 和 T790M。奥希替尼，第三代 EGFR 酪氨酸激酶抑制剂（EGFR-TKI），对 NSCLC 患者的 EGFR-T790M 突变具有治疗功效；然而，奥希替尼的治疗仍可导致肺癌患者产生耐药性。因此，研究奥希替尼耐药的机制将为肺癌治疗提供新的策略。
方法
通过用奥希替尼长期孵育 H1975 细胞获得奥希替尼耐药细胞系 H1975OR。通过细胞活力和细胞生长测定评估细胞增殖能力。通过 Boyden 室测定评估细胞迁移能力。通过全基因组 RNA 测序分析差异基因表达谱。通过 Western blot 和共聚焦显微镜分析蛋白表达和定位。
结果
在这项研究中，我们建立了奥希替尼耐药的 H1975（T790M/L858R）癌细胞，命名为 H1975OR 细胞系。与 H1975 亲本细胞相比，H1975OR 细胞的细胞生长能力降低。相反，H1975OR 细胞的迁移能力增强。我们发现 H1975OR 和 H1975 亲本细胞之间增殖和迁移途径的差异基因表达谱。有趣的是，磷酸化 EGFR、PD-L1、E-钙粘蛋白和β-连环蛋白的蛋白水平降低，但 H1975OR 耐药细胞中的存活素和 N-钙粘蛋白蛋白水平升高。
结论
奥希替尼诱导 EGFR-T790M 肺癌细胞耐药时增殖和迁移的相反作用。我们认为，细胞增殖和迁移途径的差异基因和蛋白表达可能介导奥希替尼在肺癌细胞中的耐药性。了解奥希替尼增殖和迁移途径的分子耐药机制可能为 EGFR-TKIs 治疗肺癌患者提供新的靶点和策略。
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