1749. |
Rac1 and Rho contribute to the migratory and invasive phenotype associated with somatic E-cadherin mutation
Joelle Deplazes, Margit Fuchs, Sandra Rauser, Harald Genth, Ernst Lengyel, Raymonde Busch, Birgit Luber,
Technische Universitat Muenchen,
Human Molecular Genetics,
18(1749),
(2009)
Link To Paper
Materials & Methods:
Cell cultivation and transfection: Wild-type and mutant E-cadherin cDNAs were isolated previously from non-tumourous gastric mucosa or somatic gastric carcinoma of the diffuse type and were expressed stably in MDA-MB-435S carcinoma cells (ATCC, Rockville, MD, USA) (10). Vector-transfected control MDA-MB-435S cells were obtained after transfection with the b-actin promoter-based pBATEM expression vector as described before (10). The cells were grown in Dulbecco’s modified Eagle’s medium (Life Technologies, Eggenstein, Germany) supplemented with 10% fetal calf serum (PAN-Biotech, Aidenbach, Germany) and penicillin–streptomycin (50 IU/ml and 50 mg/ml; Life Technologies) at 378C and 5% CO2. The MDA-MB-435S strain evolved from the parent line MDA-MB-435, which was isolated from the pleural effusion of a female with metastatic, ductal adenocarcinoma of the breast (68). Microscopic Technique
Epifluorescence microscope Cell Type(s)
MDA-MB-435S carcinoma cells |
Cart
plate with a glass bottom (MatTek Corporation, Ashland, MA, USA) 