Strategies for the purification of laminin-10 for studies on colon cancer metastasis.
Signals from the epidermal growth factor (EGF) receptor family are thought to combine with integrin-dependent adhesion to laminins to contribute to disease progression and metastasis in cancer. To date, little is known about the mechanisms by which these signals interact. Recently, we have shown that the colon cancer cell line LIM1215 secretes and adheres to laminin-10 through multiple integrin receptors, and that EGF stimulates spreading and migration of these cells on the same substrate. Additionally laminin-10/11 has been shown by immunohistochemistry to be present at the invasive edge of moderately differentiated colon cancers. To enable detailed structure-function studies to be undertaken, it is important to be able to rapidly obtain highly purified native laminin-10 from bulk biological samples in reasonable yield. The development of a multidimensional micropurification scheme to achieve this is presented and compared with other reported methods for the purification of laminins.
Zamurs
Laura
L
The Ludwig Institute for Cancer Research, Melbourne Tumour Biology Branch, Melbourne, Victoria, Australia.
Pouliot
Normand
N
Gibson
Peter
P
Hocking
Glenn
G
Nice
Edouard
E
D045744:Cell Line, Tumor; D002853:Chromatography, Liquid / Q000379:methods; D003110:Colonic Neoplasms / Q000473:pathology*; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007797:Laminin / Q000302:isolation & purification*; D009362:Neoplasm Metastasis
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D007797:Laminin; C119013:laminin 10