학술논문
High threshold of [beta]1 integrin inhibition required to block collagen I-induced membrane type-1 matrix metalloproteinase (MT1-MMP) activation of matrix metalloproteinase 2 (MMP-2)
Document Type
Academic Journal
Source
Cancer Cell International. October 3, 2014, Vol. 14
Subject
Language
English
Abstract
Author(s): Kulrut Borrirukwanit[sup.1,2] , Prasit Pavasant[sup.3] , Tony Blick[sup.2,5] , Marc A Lafleur[sup.2] and Erik W Thompson[sup.2,4,5] Background Integrins are a family of transmembrane adhesion receptors that mediate cell-matrix and [...]
Background Matrix metalloproteinase-2 (MMP-2) is an endopeptidase that facilitates extracellular matrix remodeling and molecular regulation, and is implicated in tumor metastasis. Type I collagen (Col I) regulates the activation of MMP-2 through both transcriptional and post-transcriptional means; however gaps remain in our understanding of the involvement of collagen-binding [beta]1 integrins in collagen-stimulated MMP-2 activation. Methods Three [beta]1 integrin siRNAs were used to elucidate the involvement of [beta]1 integrins in the Col I-induced MMP-2 activation mechanism. were used to elucidate the involvement of1 integrin knockdown was analyzed by quantitative RT-PCR, Western Blot and FACS analysis. Adhesion assay and collagen gel contraction were used to test the biological effects of [beta]1 integrin abrogation. MMP-2 activation levels were monitored by gelatin zymography. Results All three [beta]1 integrin siRNAs were efficient at [beta]1 integrin knockdown and FACS analysis revealed commensurate reductions of integrins [alpha]2 and [alpha]3, which are heterodimeric partners of [beta]1, but not [alpha]V, which is not. All three [beta]1 integrin siRNAs inhibited adhesion and collagen gel contraction, however only the siRNA showing the greatest magnitude of [beta]1 knockdown inhibited Col I-induced MMP-2 activation and reduced the accompanying upregulation of MT1-MMP, suggesting a dose response threshold effect. Re-transfection with codon-swapped [beta]1 integrin overcame the reduction in MMP-2 activation induced by Col-1, confirming the [beta]1 integrin target specificity. MMP-2 activation induced by TPA or Concanavalin A (Con A) was not inhibited by [beta]1 integrin siRNA knockdown. Conclusion Together, the data reveals that strong abrogation of [beta]1 integrin is required to block MMP-2 activation induced by Col I, which may have implications for the therapeutic targeting of [beta]1 integrin. Keywords: [beta]1 Integrin, Matrix metalloproteinase, MMP-2 activation, Type I collagen, Invasion and metastasis
Background Matrix metalloproteinase-2 (MMP-2) is an endopeptidase that facilitates extracellular matrix remodeling and molecular regulation, and is implicated in tumor metastasis. Type I collagen (Col I) regulates the activation of MMP-2 through both transcriptional and post-transcriptional means; however gaps remain in our understanding of the involvement of collagen-binding [beta]1 integrins in collagen-stimulated MMP-2 activation. Methods Three [beta]1 integrin siRNAs were used to elucidate the involvement of [beta]1 integrins in the Col I-induced MMP-2 activation mechanism. were used to elucidate the involvement of1 integrin knockdown was analyzed by quantitative RT-PCR, Western Blot and FACS analysis. Adhesion assay and collagen gel contraction were used to test the biological effects of [beta]1 integrin abrogation. MMP-2 activation levels were monitored by gelatin zymography. Results All three [beta]1 integrin siRNAs were efficient at [beta]1 integrin knockdown and FACS analysis revealed commensurate reductions of integrins [alpha]2 and [alpha]3, which are heterodimeric partners of [beta]1, but not [alpha]V, which is not. All three [beta]1 integrin siRNAs inhibited adhesion and collagen gel contraction, however only the siRNA showing the greatest magnitude of [beta]1 knockdown inhibited Col I-induced MMP-2 activation and reduced the accompanying upregulation of MT1-MMP, suggesting a dose response threshold effect. Re-transfection with codon-swapped [beta]1 integrin overcame the reduction in MMP-2 activation induced by Col-1, confirming the [beta]1 integrin target specificity. MMP-2 activation induced by TPA or Concanavalin A (Con A) was not inhibited by [beta]1 integrin siRNA knockdown. Conclusion Together, the data reveals that strong abrogation of [beta]1 integrin is required to block MMP-2 activation induced by Col I, which may have implications for the therapeutic targeting of [beta]1 integrin. Keywords: [beta]1 Integrin, Matrix metalloproteinase, MMP-2 activation, Type I collagen, Invasion and metastasis