PDF file - 117KB, Supplementary figure 5: Rescue of EGFR inhibition by active Src in Fhit siRNA-treated cells. (a) Western blot analysis of phospho-Src, total Src, Fhit and Slug levels in HBE4-E6/E7 cells transduced with control or Src. (b) Analysis by Western blotting and zymography of the effect of Src overexpression on vimentin and MMP-9 levels in Fhit siRNA transfectants treated with DMSO or gefitinib.
ARTICLE ABSTRACT
In many cancers, including lung carcinomas, Fragile histidine triad (Fhit) is frequently decreased or lost. Fhit status has recently been shown to be associated with elevated in vitro and in vivo invasiveness in lung cancer. Tumor cell invasion is facilitated by epithelial–mesenchymal transition (EMT), a process by which tumor cells lose their epithelial features to acquire a mesenchymal cell-like phenotype. In this study, the mechanism underlying Fhit-regulated EMT was deciphered. Using Slug knockdown, pharmacologic inhibitors PD98059, PP1, and gefitinib as well as an anti-EGFR antibody, it was demonstrated that Fhit silencing in bronchial cells induced overexpression of two primary EMT-associated targets, MMP-9 and vimentin, to regulate cell invasion dependent on an EGFR/Src/ERK/Slug signaling pathway. Moreover, ectopic expression of Fhit in Fhit-deficient lung cancer cells downregulated this pathway. Finally, an inverse correlation was observed between Fhit and phospho-EGFR levels in a cohort of human squamous cell lung carcinoma specimens. These results demonstrate a Fhit-dependent mechanism in the control of EMT-regulated EGFR signaling.Implications: This study adds new insight into the regulatory mechanism of EMT, a process known to increase resistance to conventional and targeted therapies in lung cancer. Mol Cancer Res; 12(5); 775–83. ©2014 AACR.
