Functional live-cell imaging demonstrates that beta1-integrin promotes type IV collagen degradation by breast and prostate cancer cells.
The ability of tumor cells to adhere to, migrate on, and remodel extracellular matrices is mediated by cell surface receptors such as beta1-integrins. Here we conducted functional live-cell imaging in real time to investigate the effects of modulating beta1-integrin expression and function on proteolytic remodeling of the extracellular matrix. Human breast and prostate cancer cells were grown on reconstituted basement membrane containing a quenched fluorescent form of collagen IV. Generation of cleavage products and the resulting increases in fluorescence were imaged and quantified. Decreases in the expression and activity of beta1-integrin reduced digestion of quenched fluorescent-collagen IV by the breast and prostate cancer cells and correspondingly their invasion through and migration on reconstituted basement membrane. Decreased extracellular matrix degradation also was associated with changes in the constituents of proteolytic pathways: decreases in secretion of the cysteine protease cathepsin B, the matrix metalloproteinase (MMP)-13, and tissue inhibitors of metalloproteinases (TIMP)-1 and 2; a decrease in expression of MMP-14 or membrane type 1 MMP; and an increase in secretion of TIMP-3. This is the first study to demonstrate through functional live-cell imaging that downregulation of beta1-integrin expression and function reduces proteolysis of collagen IV by breast and prostate cancer cells.
Sameni
Mansoureh
M
Department of Pharmacology and Barbara Ann Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA.
Dosescu
Julie
J
Yamada
Kenneth M
KM
Sloane
Bonnie F
BF
Cavallo-Medved
Dora
D
D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002401:Cathepsin B / Q000032:analysis / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D024141:Collagen Type IV / Q000378:metabolism*; D017077:Culture Media, Conditioned / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D003952:Diagnostic Imaging; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000378:metabolism*; D008297:Male; D009361:Neoplasm Invasiveness / Q000503:physiopathology; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology
D024141:Collagen Type IV; D017077:Culture Media, Conditioned; D019012:Integrin beta1; D004247:DNA; D002401:Cathepsin B