Differential modulation of human melanoma cell metalloproteinase expression by alpha2beta1 integrin and CD44 triple-helical ligands derived from type IV collagen.
Tumor cell binding to components of the basement membrane is well known to trigger intracellular signaling pathways. Signaling ultimately results in the modulation of gene expression, facilitating metastasis. Type IV collagen is the major structural component of the basement membrane and is known to be a polyvalent ligand, possessing sequences bound by the alpha1beta1, alpha2beta1, and alpha3beta1 integrins, as well as cell surface proteoglycan receptors, such as CD44/chondroitin sulfate proteoglycan (CSPG). The role of alpha2beta1 integrin and CD44/CSPG receptor binding on human melanoma cell activation has been evaluated herein using triple-helical peptide ligands incorporating the alpha1(IV)382-393 and alpha1(IV)1263-1277 sequences, respectively. Gene expression and protein production of matrix metalloproteinases-1 (MMP-1), -2, -3, -13, and -14 were modulated with the alpha2beta1-specific sequence, whereas the CD44-specific sequence yielded significant stimulation of MMP-8 and lower levels of modulation of MMP-1, -2, -13, and -14. Analysis of enzyme activity confirmed different melanoma cell proteolytic potentials based on engagement of either the alpha2beta1 integrin or CD44/CSPG. These results are indicative of specific activation events that tumor cells undergo upon binding to select regions of basement membrane collagen. Based on the present study, triple-helical peptide ligands provide a general approach for monitoring the regulation of proteolysis in cellular systems.
Department of Chemistry and Biochemistry, Florida Atlantic University, Boca Raton 33431-0991, USA.
Diane
D
Baronas-Lowell
Janelle L
JL
Lauer-Fields
Jeffrey A
JA
Borgia
Gian Franco
GF
Sferrazza
Mohammad
M
Al-Ghoul
Dmitriy
D
Minond
Gregg B
GB
Fields
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000502:physiology; D001483:Base Sequence; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D024141:Collagen Type IV / Q000737:chemistry / Q000494:pharmacology*; D017931:DNA Primers; D006801:Humans; D018960:Hyaluronan Receptors / Q000502:physiology*; D008024:Ligands; D008545:Melanoma; D045726:Metalloproteases / Q000187:drug effects / Q000235:genetics*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D015703:Antigens, CD; D024141:Collagen Type IV; D017931:DNA Primers; D018960:Hyaluronan Receptors; D008024:Ligands; D010446:Peptide Fragments; D012333:RNA, Messenger; D045726:Metalloproteases