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Estudamos inducao da atividade de MMP pelo peptideo da laminina a1 AG73 em linhagem celular (CAC2) de carcinoma adenoide cistico. CAC2 cultivadas em laminina-111 com AG73 geraram espacos pseudocisticos. Inibidor de MMP diminuiu tais espacos, sugerindo acao de MMPs. CAC2 crescidas sobre AG73 mostraram aumento dose-dependente de MMP9. RNAi para MMP9 diminuiu remodelacao em cultura 3D. Buscamos receptores de AG73 ligados a atividade de MMP9. CAC2 crescidas sobre AG73 exibiram colocalizacao de sindecan-1 e integrina b1. RNAi para sindecan-1 ou para integrina b1 geraram, isolados, reducao na adesao a AG73 e nas atividades de remodelacao e de protease. Duplo RNAi estudou a cooperacao entre os receptores e promoveu diminuicao na adesao a AG73 e na atividade de MMP. Distincao de receptores foi feita por cromatografia de afinidade e espectrometria de massa, atraves de colunas de afinidade com AG73 acoplado, que resultou em possiveis receptores, como integrinas b1 e aV. Sugerimos que AG73 regula adesao e secrecao de MMP em celulas CAC2 atraves de sindecan-1 e integrina b1.%%%%We studied induction of MMP activity by b1-laminin peptide AG73 in adenoid cystic carcinoma cell line (CAC2). Cells grown inside AG73-enriched laminin-111 exhibited pseudocystics spaces. MMP inhibitor decreased those spaces, suggesting MMPs action. Cells grown on AG73 showed a dose-dependent increase of MMP9 secretion. MMP9 siRNAi decreased remodeling in 3D culture. We searched for AG73 receptors regulating MMP9 activity. CAC2 grown on AG73 exhibited colocalization of syndecan-1 and b1 integrin. Syndecan-1 siRNA or siRNA b1 integrin showed reduction in adhesion to AG73 and in remodeling and protease activities. Double-knockdown explored syndecan-1 and 1 integrin cooperation and showed decrease in adhesion to AG73 and in MMP activity. Receptors characterization was made by affinity chromatography followed by mass spectrometry through AG73-affinity columns and showed putative receptors, like b1 and aV integrins. We suggest that AG73 peptide regulates adhesion and MMP secretion in CAC2 cells through syndecan-1 and b1 integrin.

Adesão e atividade de protease são reguladas pelo peptídeo derivado da laminina AG73, sindecan-1 e integrina 1 em linhagem celular derivada de carcinoma adenóide cístico