Effect of forced E-cadherin expression on adhesion and proliferation of human breast carcinoma cells.
- Author:
Li-Juan YANG
1
;
Yu-Qin LIU
;
Bei GU
;
Xiao-Cui BIAN
;
Hai-Liang FENG
;
Zhen-Li YANG
;
Yan-Yan LIU
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Breast Neoplasms; metabolism; pathology; Cadherins; genetics; metabolism; physiology; Cell Adhesion; Cell Line, Tumor; Cell Proliferation; Cyclin D1; metabolism; Female; Genetic Vectors; Humans; Plasmids; Transfection; beta Catenin; metabolism
- From: Chinese Journal of Pathology 2010;39(12):842-847
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the role that E-cadherin (E-cad) plays on cell adhesion and proliferation of human breast carcinoma.
METHODSE-cad expression vector was transfected into an E-cad-negative human breast carcinoma MDA-MB-231 cells. G418 was used to screen positive clones. E-cad, β-catenin (β-cat) and cyclin D1 expressions of these clones were confirmed by Western blot. Their cell-cell and cell-matrix adhesion abilities were detected. E-cad/β-catenin interaction was confirmed by immunoprecipitation. Cell proliferation was evaluated by MTT. Cell apoptosis was analyzed by flow cytometry. Direct two-step immunocytochemistry was used to detect the localization of β-cat.
RESULTE-cad(+) cell strains Ecad-231-7 and Ecad-231-9 were established. When cultured in ultra-low-binding dishes Ecad-231 cells grow in suspension while Ecad-231-7 and Ecad-231-9 cells grow in large clamps. When co-cultured with HCT116 cells, the average adhesion rates at 30 min are 39.0%, 60.0% and 59.5% for MDA-MB-231, Ecad-231-7 and Ecad-231-9 respectively. The average detachment rates by EDTA for 5 min are 37.4%, 4.2% and 7.4% respectively. So E-cad expression enhanced hemotypic and heterotypic cell-cell adhesion and cell-matrix adhesion. Forced exogenously expressed E-cad could combine with endogenous β-cat, whereas down stream cyclin D1 expression was significantly decreased, as evidenced by Western blot. The rates of cell apoptosis of MDA-MB-231, Ecad-231-7 and Ecad-231-9 were 1.8%, 2.0% and 2.1%. Expression of E-cad had no obvious effect on the apoptosis of tumor cells with regular culture. β-cat increased in the cytoplasma.
CONCLUSIONSTwo monoclonal tumor cell strains (Ecad-231-7 and Ecad-231-9) stably expressing E-cad were successfully established. E-cad could enhance adhesion and inhibit proliferation of human breast carcinoma cells through a pathway involving β-cat and cyclin D1.