title: Integrin β1-mediated acquired gefitinib resistance in non-small cell lung cancer cells occurs via the phosphoinositide 3-kinase-dependent pathway.
abstract: The present study aimed to explore the role of integrin β1 and the relevant signaling pathways in acquired gefitinib resistance in non-small cell lung cancer (NSCLC). The inhibitory effects of gefitinib, with or without LY294002, on cellular proliferation were evaluated by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide assay. Cell cycle progression and apoptosis were analyzed by flow cytometry, while western blotting was used to evaluate the expression of EGFR, phosphorylated (phospho)-EGFR, protein kinase B (Akt), phospho-Akt, extracellular signal-regulated kinase (Erk) and phospho-Erk. The gene expression profiles of PC9 and PC9/G cells were determined by DNA microarray. Integrin β1 was knocked down in PC9/G cells by transiently transfected short interfering RNA (siRNA). A scrambled siRNA sequence was used as a control. Apoptosis of transfected cells was determined by Annexin V-phycoerythrin-Cy5/propidium iodide staining. Sequencing products were amplified by nested PCR. The resistant index of PC9/G cells to gefitinib was ~138- to 256-fold higher than that of PC9 cells, and this resistance was accompanied by significant increase in integrin β1 expression in PC9/G cells. Knockdown of integrin β1 with short hairpin RNA in PC9/G cells markedly inhibited proliferation and enhanced apoptosis in response to gefitinib, restoring the sensitivity of PC9/G cells gefitinib. Phosphoinositide 3-kinase (PI3K)/Akt activation was observed in PC9/G cells in the presence of gefitinib and the sensitivity of PC9/G cells to gefitinib was also able to be restored by PI3K/Akt pathway inhibitor LY294002. Finally, knockdown of integrin β1 significantly reduced the levels of phospho-Akt. These findings suggest that integrin β1 signaling via the PI3K/Akt pathway may be a significant mechanism underlying gefitinib resistance, and may potentially present an alternative therapeutic target for the treatment of NSCLC unresponsive to EGFR inhibitors.
authors: Department of Oncology, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai 200433, P.R. China; Department of Oncology, School of Medicine, Soochow University, Suzhou 215007, P.R. China.
authors: Qin-Fang
authors: QF
authors: Deng
authors: Central Laboratory, Shanghai Pulmonary Hospital, Tongji University, Shanghai 200433, P.R. China.
authors: B O
authors: BO
authors: Su
authors: Central Laboratory, Shanghai Pulmonary Hospital, Tongji University, Shanghai 200433, P.R. China.
authors: Yin-Min
authors: YM
authors: Zhao
authors: Central Laboratory, Shanghai Pulmonary Hospital, Tongji University, Shanghai 200433, P.R. China.
authors: Liang
authors: L
authors: Tang
authors: Central Laboratory, Shanghai Pulmonary Hospital, Tongji University, Shanghai 200433, P.R. China.
authors: Jie
authors: J
authors: Zhang
authors: Department of Oncology, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai 200433, P.R. China.
authors: Cai-Cun
authors: CC
authors: Zhou
keywords: drug resistance; epidermal growth factor receptor inhibition; integrin; tyrosine kinase inhibitor