Cancer Drug Resistance Laboratory, Department of Life Science, National Institute of Technology Rourkela, Odisha 769008, India.
Cancer Drug Resistance Laboratory, Department of Life Science, National Institute of Technology Rourkela, Odisha 769008, India.
Life Sci. 2021 Jul 15;277:119608. doi: 10.1016/j.lfs.2021.119608. Epub 2021 May 11.
The first-generation epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI), gefitinib, continues to be a primary treatment option for lung cancer patients. However, acquisition of resistance to gefitinib is a major obstacle in lung cancer treatment and its cause is poorly understood. The present study aimed to implicate the role of SOX9-β-catenin in developed resistance to gefitinib through epithelial to mesenchymal transition (EMT) in lung cancer in vitro and ex vivo.
Expression effect of SOX9 on survivability of lung cancer patients was demonstrated through online available Kaplan-Meier Plotter data base. Then, cell viability assay, colony forming assay, cell migration and invasion assays, flow cytometry, drug efflux assay, qRT-PCR, and western blotting were conducted to confirmed the role of SOX9 in gefitinib resistance in lung cancer cells. Dual-luciferase assay established the regulatory relation between SOX9 and β-catenin. Multicellular spheroid assay further explored that down regulation of SOX9 could reverse gefitinib resistance ex vivo.
Kaplan-Meier method correlated the higher expression of SOX9 and β-catenin with poor overall survival of lung cancer patients. Upregulation of SOX9 was associated gefitinib resistance with increased cell proliferation, migration and invasion, single-cell colony-forming ability, reduced apoptosis, and gefitinib intake in lung cancer cells. Moreover, upregulated SOX9 promoted EMT via targeting β-catenin and knockdown of SOX9 reversed the resistance and EMT phenotype. Similarly, we found that multicellular spheroid of gefitinib resistant cells showed larger surface area with more dispersion and viability of cells, while SOX9 knockdown abolished these induced properties ex vivo.
SOX9 expression could provide an innovative perspective as biomarker to understand the EGFR-TKIs resistance in lung cancer.
第一代表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKI）吉非替尼仍是肺癌患者的主要治疗选择。然而，对吉非替尼的获得性耐药是肺癌治疗中的一个主要障碍，其原因尚不清楚。本研究旨在通过体外和体内上皮间质转化（EMT）探讨 SOX9-β-catenin 在肺癌对吉非替尼耐药中的作用。
通过在线可用的 Kaplan-Meier Plotter 数据库证明 SOX9 表达对肺癌患者生存率的影响。然后，通过细胞活力测定、集落形成测定、细胞迁移和侵袭测定、流式细胞术、药物外排测定、qRT-PCR 和 Western blot 验证 SOX9 在肺癌细胞对吉非替尼耐药中的作用。双荧光素酶报告基因实验确定了 SOX9 与 β-catenin 的调控关系。多细胞球体实验进一步探索了下调 SOX9 可在体内逆转吉非替尼耐药。
Kaplan-Meier 法将 SOX9 和 β-catenin 的高表达与肺癌患者的总生存率差相关联。SOX9 的上调与肺癌细胞中吉非替尼耐药相关，表现为细胞增殖、迁移和侵袭增加，单细胞集落形成能力增强，细胞凋亡减少，吉非替尼摄取减少。此外，上调 SOX9 通过靶向 β-catenin 促进 EMT，而 SOX9 的敲低则逆转了耐药和 EMT 表型。同样，我们发现吉非替尼耐药细胞的多细胞球体显示出更大的表面积，细胞的分散性和活力更高，而 SOX9 的敲低在体内消除了这些诱导特性。
SOX9 的表达可以提供一个创新的视角，作为生物标志物来理解肺癌中 EGFR-TKIs 的耐药性。
