title: Differential susceptibility of osteosarcoma cells and primary osteoblasts to cell detachment caused by snake venom metalloproteinase protein.
abstract: The interaction of extracellular matrix with cells plays a key role in the regulation of cell adhesion, migration, proliferation as well as differentiation. Transformed cells express a different profile of adhesion molecules, which may mediate metastasis under specific matrix microenvironment. We here found that ROS 17/2.8 osteosarcoma cells and osteoblasts have different expression of alpha5 integrin, executing different fibronectin fibrillogenesis. As compared with ROS 17/2.8 cells, osteoblasts have higher expression of fibronectin, collagen, alpha5, beta1, alpha2 integrins and focal adhesion kinase as examined by immunostaining and flow cytometry. Crovidisin, a PIII snake venom metalloproteinase (SVMP) purified from venom of Crotalus viridis, exhibits collagen-binding activity and matrix metalloproteinase activity. Crovidisin selectively caused the detachment of ROS 17/2.8 osteosarcoma cells but not of primary cultured osteoblasts. On the other hand, triflavin, an RGD-dependent disintegrin purified from venom of Trimeresurus flavoviridis, did not cause the detachment of both osteoblasts and ROS 17/2.8 cells. Although ROS 17/2.8 cells detached from substratum after crovidisin treatment for 24 h, the loss of mitochondrial membrane potential was not observed unless a prolonged treatment for longer than 36 h. These results suggest that cultured primary rat osteoblasts and ROS 17/2.8 osteosarcoma cells possess different expression of integrins and matrix environment, and ROS 17/2.8 is much more susceptible to be detached by crovidisin. The matrix degradation by crovidisin may be responsible for the preferential detachment of ROS 17/2.8 osteosarcoma cells.
authors.affiliation: Department of Pharmacology, College of Medicine, National Taiwan University, No. 1, Sec. 1 Jen-Ai Road, Taipei 100, Taiwan.
authors.forename: Chih-Hsin
authors.initials: CH
authors.lastname: Tang
authors.forename: Rong-Sen
authors.initials: RS
authors.lastname: Yang
authors.forename: Chao-Zong
authors.initials: CZ
authors.lastname: Liu
authors.forename: Tur-Fu
authors.initials: TF
authors.lastname: Huang
authors.forename: Wen-Mei
authors.initials: WM
authors.lastname: Fu
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D003094:Collagen / Q000378:metabolism; D003435:Crotalid Venoms / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism; D016023:Integrins / Q000378:metabolism; D045726:Metalloproteases / Q000494:pharmacology*; D010006:Osteoblasts / Q000378:metabolism*; D012516:Osteosarcoma / Q000378:metabolism* / Q000473:pathology; D010455:Peptides / Q000378:metabolism; D051381:Rats; D030162:Reptilian Proteins; D014407:Tumor Cells, Cultured
chemical_list: D002352:Carrier Proteins; D003435:Crotalid Venoms; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D016023:Integrins; D010455:Peptides; D030162:Reptilian Proteins; C070025:triflavin; D003094:Collagen; D045726:Metalloproteases; C103975:crovidisin protein, Crotalus viridis