Antitumour efficacy of MEK inhibitors in human lung cancer cells and their derivatives with acquired resistance to different tyrosine kinase inhibitors

• Published in: British journal of cancer Vol. 105; no. 3; pp. 382 - 392
• Main Authors: Morgillo, F, Cascone, T, D'Aiuto, E, Martinelli, E, Troiani, T, Saintigny, P, De Palma, R, Heymach, J V, Berrino, L, Tuccillo, C, Ciardiello, F
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 26.07.2011; Nature Publishing Group
• Subjects: 631/67/1059/2326; 631/92/436/2388; 692/699/67/1612; 692/700/565/1436; Adenocarcinoma - drug therapy; Adenocarcinoma of Lung; Animal tumors. Experimental tumors; Animals; Antineoplastic agents; Benzenesulfonates - pharmacology; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Cancer Research; Cell Line, Tumor; Cell Proliferation - drug effects; Chemotherapy; Drug Resistance; Drug Resistance, Neoplasm; Epidemiology; Epithelial-Mesenchymal Transition - drug effects; Erlotinib Hydrochloride; Experimental tumors, general aspects; Gefitinib; Gene Expression Profiling; Humans; Lung cancer; Lung Neoplasms - drug therapy; MAP Kinase Kinase Kinases - antagonists & inhibitors; Medical sciences; Mice; Mice, Nude; Molecular Medicine; Niacinamide - analogs & derivatives; Oncology; Pharmacology. Drug treatments; Phenylurea Compounds; Piperidines - pharmacology; Protein Kinase Inhibitors - therapeutic use; Pyridines - pharmacology; Quinazolines - pharmacology; Sorafenib; Translational Therapeutics; Tumors; Xenograft Model Antitumor Assays; MEK; gefitinib; vandetanib; EMT; erlotinib; acquired resistance; Antineoplastic agent; Vandetanib; Acquired; Quinazoline derivatives; Lung cancer; Bronchopulmonary adenocarcinoma; Cancerology; Established cell line; Bronchus disease; Antiangiogenic agent; Gefitinib; Protein-tyrosine kinase; Human; Lung disease; Treatment resistance; Enzyme; Respiratory disease; Tyrosine kinase inhibitor; Transferases; Treatment efficiency; Rodentia; Enzyme inhibitor; Mitogen-activated protein kinase; Malignant tumor; In vitro; In vivo; Vertebrata; Chemotherapy; Mammalia; Treatment; Erlotinib; Mouse; Animal; Multikinase inhibitor; Cancer
• ISSN: 0007-0920; 1532-1827
• DOI: 10.1038/bjc.2011.244

Background: To study the molecular mechanisms regulating cancer cell resistance to four different tyrosine kinase inhibitors (TKIs): erlotinib, gefitinib, vandetanib and sorafenib. Methods: An in vitro model of acquired resistance to these TKIs was developed by continuously treating the human lung adenocarcinoma cell line CALU-3 with escalating doses of each drug. Transcriptional profiling was performed with Agilent whole genome microarrays. Western blot analysis, enzyme-linked immunosorbent (ELISA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation, migration, invasion and anchorage-independent colony growth assays were conducted in vitro and experiments with established xenografts in athymic nude mice were performed in vivo in parental (P) and TKI-resistant (R) CALU-3 cell lines. Results: As compared with P-CALU-3 cells, in TKI-R CALU-3 cell lines a significant increase in the expression of activated, phosphorylated MET, IGF-1R, AKT, MEK, MAPK and of survivin was observed. Downregulation of E-cadherin and amphiregulin mRNAs and upregulation of vimentin, VE-cadherin, HIF-1 α and vascular endothelial growth factor receptor-1 mRNAs were observed in all four TKI-R CALU-3 cell lines. All four TKI-R CALU-3 cells showed increased invasion, migration and anchorage-independent growth. Together, these data suggest epithelial to mesenchymal transition (EMT) in TKI-R CALU-3 cells. Treatment with several agents that target AKT, MET or IGF-1R did not affect TKI-R CALU-3 cell proliferation. In contrast, treatment with MSC19363669B and selumetinib, two selective MEK inhibitors, caused inhibition of cell proliferation, invasion, migration, anchorage-independent growth in vitro and of tumour growth in vivo of all four TKI-R CALU-3 cell lines. Conclusion: These data suggest that resistance to four different TKIs is characterised by EMT, which is MEK-inhibitor sensitive in human CALU-3 lung adenocarcinoma.