Technical Reference #1749

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