ABSTRACT
We have investigated the role of vascular-endothelial (VE)-cadherin in melanoma and breast cancer metastasis. We found that VE-cadherin is expressed in highly aggressive melanoma and breast cancer cell lines. Remarkably, inactivation of VE-cadherin triggered a significant loss of malignant traits (proliferation, adhesion, invasion and transendothelial migration) in melanoma and breast cancer cells. These effects, except transendothelial migration, were induced by the VE-cadherin RGD motifs. Co-immunoprecipitation experiments demonstrated an interaction between VE-cadherin and α2ß1 integrin, with the RGD motifs found to directly affect ß1 integrin activation. VE-cadherin-mediated integrin signaling occurred through specific activation of SRC, ERK and JNK, including AKT in melanoma. Knocking down VE-cadherin suppressed lung colonization capacity of melanoma or breast cancer cells inoculated in mice, while pre-incubation with VE-cadherin RGD peptides promoted lung metastasis for both cancer types. Finally, an in silico study revealed the association of high VE-cadherin expression with poor survival in a subset of melanoma patients and breast cancer patients showing low CD34 expression. These findings support a general role for VE-cadherin and other RGD cadherins as critical regulators of lung and liver metastasis in multiple solid tumours. These results pave the way for cadherin-specific RGD targeted therapies to control disseminated metastasis in multiple cancers.
Subject(s)
Antigens, CD/metabolism , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Cadherins/metabolism , Melanoma/metabolism , Melanoma/pathology , Oligopeptides , Protein Interaction Domains and Motifs , Animals , Antigens, CD/chemistry , Antigens, CD/genetics , Antineoplastic Agents/pharmacology , Breast Neoplasms/drug therapy , Cadherins/antagonists & inhibitors , Cadherins/chemistry , Cadherins/genetics , Cell Adhesion , Cell Line, Tumor , Cell Movement , Cell Proliferation , Disease Models, Animal , Female , Gene Expression , Heterografts , Humans , Integrins/metabolism , Melanoma/drug therapy , Melanoma/mortality , Mice , Models, Biological , Molecular Targeted Therapy , Neoplasm Metastasis , Prognosis , Protein Binding , Protein Interaction Maps , Signal TransductionABSTRACT
Stimulation by chemokines of integrin α4ß1-dependent T-lymphocyte adhesion is a crucial step for lymphocyte trafficking. The adaptor Vav1 is required for chemokine-activated T-cell adhesion mediated by α4ß1. Conceivably, proteins associating with Vav1 could potentially modulate this adhesion. Correlating with activation by the chemokine CXCL12 of T-lymphocyte attachment to α4ß1 ligands, a transient stimulation in the association of Vav1 with SLP-76, Pyk2, and ADAP was observed. Using T-cells depleted for SLP-76, ADAP, or Pyk2, or expressing Pyk2 kinase-inactive forms, we show that SLP-76 and ADAP stimulate chemokine-activated, α4ß1-mediated adhesion, whereas Pyk2 opposes T-cell attachment. While CXCL12-promoted generation of high-affinity α4ß1 is independent of SLP-76, ADAP, and Pyk2, the strength of α4ß1-VCAM-1 interaction and cell spreading on VCAM-1 are targets of regulation by these three proteins. GTPase assays, expression of activated or dominant-negative Rac1, or combined ADAP and Pyk2 silencing indicated that Rac1 activation by CXCL12 is a common mediator response in SLP-76-, ADAP-, and Pyk2-regulated cell adhesion involving α4ß1. Our data strongly suggest that chemokine-stimulated associations between Vav1, SLP-76, and ADAP facilitate Rac1 activation and α4ß1-mediated adhesion, whereas Pyk2 opposes this adhesion by limiting Rac1 activation.
