The role of tyrosine kinase Etk/Bmx in EGF-induced apoptosis of MDA-MB-468 breast cancer cells.
Etk/Bmx, a member of the Tec family of tyrosine kinases, mediates various signaling pathways and confers several cellular functions. In the present study, we have explored the functional role of Etk in mediating EGF-induced apoptosis, using MDA-MB-468 cell line as a model. We first demonstrated that EGF treatment induces Etk tyrosine phosphorylation in both HeLa and MDA-MB-468 cells. Overexpression of Etk by recombinant adenovirus in MDA-MB-468 cells potentiates the extent of EGF-induced cell apoptosis. The observed Etk-enhanced MDA-MB-468 cell apoptosis is associated with the Stat1 activation, as demonstrated by electrophoresis mobility shift assays and reporter gene assays. By contrast, a kinase domain deletion mutant EtkDeltaK, functioning as a dominant-negative mutant, ameliorates EGF-induced Stat1 activation and apoptosis in MDA-MB-468 cells. To explore whether the activated Etk alone is sufficient for inducing apoptosis, a conditionally activated Etk (DeltaEtk-ER), a chimeric fusion protein of PH domain-truncated Etk and ligand-binding domain of estrogen receptor, was introduced into MDA-MB-468 cells. Upon beta-estradiol ligand activation, the DeltaEtk-ER could stimulate Stat1 activity and confer cell apoptosis independent of EGF treatment. Taken together, our findings indicate that Etk is a downstream signaling molecule of EGF receptor and suggest that Etk activation is essential for transducing the EGF-induced apoptotic signaling.
Chen
Kai-Yun
KY
Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, ROC.
Huang
Li-Ming
LM
Kung
Hsing-Jien
HJ
Ann
David K
DK
Shih
Hsiu-Ming
HM
D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001483:Base Sequence; D001943:Breast Neoplasms; D045744:Cell Line, Tumor; D017931:DNA Primers; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005260:Female; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D016296:Mutagenesis; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
D017931:DNA Primers; D008565:Membrane Proteins; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; C492367:BMX protein, human; D011505:Protein-Tyrosine Kinases