seryl-isoleucyl-lysyl-valyl-alanyl-valinamide and Salivary-Gland-Neoplasms

Adenoid cystic carcinoma is a frequently occurring malignant salivary gland neoplasm. We studied the induction of protease activity by the laminin-derived peptide, SIKVAV, in cells (CAC2) derived from this neoplasm. Laminin alpha1 and matrix metalloproteinases (MMPs) 2 and 9 were immunolocalized in adenoid cystic carcinoma cells in vivo and in vitro. CAC2 cells cultured on SIKVAV showed a dose-dependent increase of MMP9 as detected by zymography and colocalization of alpha3 and alpha6 integrins. Small interfering RNA (siRNA) knockdown of integrin expression in CAC2 cells resulted in decreased adhesion to the peptide. SIKVAV affinity chromatography and immunoblot analysis showed that alpha3, alpha6, and beta1 integrins were eluted from the SIKVAV column, which was confirmed by mass spectrometry and a solid-phase binding assay. Small interfering RNA experiments also showed that these integrins, through extracellular signal-regulated kinase (ERK) 1/2 signaling, regulate MMP secretion induced by SIKVAV in CAC2 cells. We propose that SIKVAV increases protease activity of a human salivary gland adenoid cystic carcinoma cell line through alpha3beta1 and alpha6beta1 integrins and the ERK 1/2 signaling pathway.

Topics: Carcinoma, Adenoid Cystic; Humans; Integrin alpha6; Integrin beta1; Integrins; Laminin; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinases; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Oligopeptides; Salivary Gland Neoplasms; Signal Transduction; Tumor Cells, Cultured

We have demonstrated that the basement membrane regulates the myoepithelioma. We are now studying the effect of laminin, a basement membrane protein, in the morphology of a cell line (M1) derived from human salivary gland plasmacytoid myoepithelioma. These cells were grown inside a three-dimensional preparation of laminin-1. Phenotype differences were assessed by light and transmission electron microscopy. In addition, we analysed the effect of a molecular domain of laminin-1, the peptide SIKVAV, on M1 cells. This peptide was chosen because it is effective in cell proliferation and differentiation. M1 cells grown inside laminin-1 were mostly plasmacytoid-like, while cells treated by SIKVAV showed light and electron microscopic features of typical plasmacytoid cells. This peptide also modulated smooth-muscle actin expression in M1 cells. We demonstrated that laminin-1 and its derived peptide SIKVAV morphoregulates myoepithelioma cells in culture.

Topics: Cell Line, Tumor; Humans; Immunohistochemistry; Laminin; Microscopy, Electron; Muscle, Smooth; Myoepithelioma; Oligopeptides; Salivary Gland Neoplasms

In a previous paper, we demonstrated that laminin-1 and its derived peptide SIKVAV modulates the morphology of an adenoid cystic carcinoma cell line (CAC2 cells). Light microscopy of CAC2 cells grown in three-dimensional preparations of SIKVAV-enriched laminin-1 showed the presence of pseudocystic spaces. Pseudocysts are hallmarks of adenoid cystic carcinoma in vivo. We hypothesized that these pseudocystic spaces could be due to the protease-inducing/activating role of SIKVAV. Thus, we studied the presence of matrix metalloproteinases (MMPs) in CAC2 cells treated either by laminin-1 or by SIKVAV-enriched laminin-1. Immunohistochemistry and zymography suggested that SIKVAV enhanced the secretion of MMP-2 and MMP-9 in CAC2 cells. We propose that SIKVAV induces pseudocystic formation probably through the secretion of MMPs 2 and 9.

Topics: Carcinoma, Adenoid Cystic; Cell Line, Tumor; Culture Media; Cytoplasm; Extracellular Space; Humans; Immunohistochemistry; Laminin; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinases; Oligopeptides; Salivary Gland Neoplasms

We have previously demonstrated that laminin modulates the expression of adhesion molecules in an adenoid cystic carcinoma cell line (CAC2 cells). We are currently studying whether laminin can induce modifications in the overall morphology of CAC2 cells. These cells were grown in a three-dimensional preparation of laminin-1. Phenotype differences were assessed by light and transmission electron microscopy. CAC2 cells grown inside laminin-1 formed ductlike and pseudocystic structures. Based on our findings we suggest that laminin is a key regulator of tubular and pseudocystic patterns of adenoid cystic carcinoma. We also analyzed the effect of a molecular domain of laminin-1, the peptide SIKVAV (Ser-Ile-Lys-Val-Ala-Val) on CAC2 cells. This peptide was chosen because it is effective in cell proliferation and differentiation, and because it has never been tested before in salivary gland neoplasms. When CAC2 cells were grown inside SIKVAV-enriched laminin-1, only pseudocystic structures were observed. Since no ductlike structures were observed in samples treated with SIKVAV, we may assume that this peptide is at least one of the molecular domains of laminin responsible for the pseudocystic pattern observed in adenoid cystic carcinoma. Function disturbing experiments strongly suggested that the integrin alpha3beta1 play a role in the effect of laminin on CAC2 cells.

Topics: Antibodies, Blocking; Carcinoma, Adenoid Cystic; Cell Adhesion; Fluorescent Antibody Technique, Indirect; Humans; Integrins; Laminin; Oligopeptides; Organoids; Salivary Gland Neoplasms; Tumor Cells, Cultured