Effect of Cetuximab and Small Interfering RNA Combination Treatment in NSCLC Cell Lines with Wild Type and Use of as a Possible Biomarker for Treatment Responsiveness.

Background: The epidermal growth factor receptor (EGFR) is a therapeutic target for patients with non-small cell lung cancer (NSCLC). Cetuximab is an anti-EGFR monoclonal antibody that inhibits EGFR signaling and proliferation of colorectal cancer and head and neck cancers. Since only few NSCLC patients benefit from cetuximab therapy, we evaluated a novel combination treatment using cetuximab and small interfering RNA (siRNA) to strongly suppress EGFR signaling and searched for a biomarker in NSCLC cell lines harboring wild-type .

Methods: Alterations in and its downstream genes in five NSCLC cell lines (A549, Lu99, 86-2, Sq19 and Ma10) were assessed through sequencing. The protein expression levels of these molecules were assessed through western blotting. The effect of combination treatment was determined through cell proliferation assay, caspase-3/7 assay, invasion assay, and migration assay.

Results: All cell lines were harboring wild-type , whereas , , and were mutated in A549 and Lu99; Lu99 and Sq19; Lu99, 86-2, Sq19 and Ma10; and A549, 86-2, and Sq19 cell lines, respectively. PTEN was not expressed in Sq19, and LKB1 was not expressed in both A549 and Sq19. TP53 was not expressed in both A549 and Lu99. The combination of cetuximab and siRNA significantly suppressed cell proliferation in 86-2, Sq19 and Ma10, which express wild-type . It induced apoptosis in A549, 86-2 and Ma10 cells, which express wild type . The combination treatment had no effect either on cell invasion nor migration in all cell lines.

Conclusion: targeted therapy using the combination of cetuximab and siRNA is effective in NSCLC cell lines harboring wild-type . Wild-type may act as a potential biomarker for response to combination treatment by the induction of apoptosis in cells with wild-type .