Antitumour effects of PLC-γ1-(SH2)2-TAT fusion proteins on EGFR/c-erbB-2-positive breast cancer cells

Abstract

Due to its pivotal role in the growth factor-mediated tumour cell migration, the adaptor protein phospholipase C-γ1 (PLC-γ1) is an appropriate target to block ultimately the spreading of EGFR/c-erbB-2-positive tumour cells, thereby minimising metastasis formation. Here, we present an approach to block PLC-γ1 activity by using protein-based PLC-γ1 inhibitors consisting of PLC-γ1 SH2 domains, which were fused to the TAT-transduction domain to ensure a high protein transduction efficiency. Two proteins were generated containing one PLC-γ1-SH2-domain (PS1-TAT) or two PLC-γ1-SH2 domains (PS2-TAT). PS2-TAT treatment of the EGFR/c-erbB-2-positive cell line MDA-HER2 resulted in a reduction of the EGF-rnediated PLC-γ1 tyrosine phosphorylation of about 30%, concomitant with a complete abrogation of the EGF-driven calcium influx. In addition to this, long-term PS2-TAT treatment both reduces the EGF-rnediated migration of about 75% combined with a markedly decreased time locomotion of single MDA-HER2 cells as well as decreases the proliferation of MDA-HER2 cells by about 50%. Due to its antitumoral capacity on EGFR/c-erbB-2-positive breast cancer cells, we conclude from our results that the protein-based PLC-γ1 inhibitor PS2-TAT may be a means for novel adjuvant antiturnour strategies to minimise metastasis formation because of the blockade of cell migration and proliferation. © 2004 Cancer Research UK.