RESUMO
Cancer cells have been known to overexpress the epidermal growth factor receptor (EGFR) and hence relevant multipletargeted therapies have been developed, with a recent clinical application of the antibodymediated inhibition of the EGFR. However, this strategy is not useful in cancer cells with mutations in KRAS; a GTPase downstream of EGFR which constitutively activates the pathway without EGF stimulation. Furthermore, mutations in EGFR also reduce the binding of monoclonal antibodies and thereby render them ineffective. In the present study, we designed a chimeric EGF protein fused to the truncated Nterminal domain fragment of Pseudomonas aeruginosa exotoxin A (EGFETA), which has ADPribosylation activity and induces apoptosis. The EGFETA protein was expressed in E. coli as a Histagged fusion. Our results showed that EGFETA significantly inhibited the proliferation of EGFRpositive A431 epidermoid carcinoma (IC50 27 ng/ml) and HN5 head and neck squamous cell carcinoma (IC50 36 ng/ml) cells. However, its effect on cancer cells with little or no EGFR expression was limited (A549IC50 1,000 ng/ml; MCF7IC50 >10,000 ng/ml). Compared to cetuximab, EGFETA was highly potent in its killing capacity of HN5 cancer cells at 1,000 ng/ml, while cetuximab had little effect at 1,000 ng/ml. Furthermore, EGFETA was just as potent in HCT116 (KRAS G13D) and SW480 (KRAS G12V) colon cancer cell lines harbouring KRAS hyperactivating mutations when compared to KRAS wildtype HT29 colon cancer cells. Finally, coincubation of EGFETA with an antiEGF antibody abrogated its effect on the EGFRpositive A431 cells. Our results show that the chimeric EGFETA toxin is extremely effective against EGFRpositive cancers and raises the potential to further develop this chimera for use in targeting EGFRpositive tumours resistant to monoclonal antibodies.