Effects of caveolin-1 on biologic behavior of laryngeal squamous cell carcinoma HEp2 cell line GU.
- Author:
Dong-hua GU
1
;
Feng TANG
;
Zhen WANG
;
Hua LI
;
Qi CHEN
;
Hong-guang ZHU
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Blotting, Western; Carcinoma, Squamous Cell; genetics; metabolism; pathology; Caveolin 1; genetics; metabolism; physiology; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Flow Cytometry; Genetic Vectors; chemistry; genetics; Humans; Laryngeal Neoplasms; genetics; metabolism; pathology; Lipids; chemistry; Microscopy, Confocal; Mitogen-Activated Protein Kinase 3; metabolism; Phosphorylation; Polymerase Chain Reaction; methods; Receptor, Epidermal Growth Factor; metabolism; Signal Transduction; physiology; Transfection; methods
- From: Chinese Journal of Oncology 2007;29(5):329-333
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effects of caveolin-1 on the biologic behavior of laryngeal squamous cell carcinoma HEp2 cell line in vitro.
METHODSEukaryotic expression vector of human caveolin-1 gene was constructed and transfected into HEp2 cells by Lipofectamine. The clones stably overexpressing caveolin-1 were identified by real-time PCR and Western blotting. Cell proliferation viability was tested by MTT assay. Anchorage-independent growth was determined by assaying colony formation in soft agar. Flow cytometry was used to assess the cell cycle and apoptosis. The relative phosphorylation level of EGFR and ERK1/2 were detected by Western blotting. Localization of caveolin-1 and EGFR were studied by laser confocal laser scanning microscopy.
RESULTSThe expression vector of caveolin-1 was constructed and three clones stably overexpressing caveolin-1 were obtained. Comparing with the parental HEp2 cells, the transfected cells exhibited a slower growth rate and formed fewer colonies in soft agar. The results of FACS analysis revealed that overexpression of caveolin-1 resulted in the cell cycle arrest at G0/G1 phase and increased the apoptotic cell fraction. EGFR was found to colocalize with caveolin-1 in transfected cells by confocal laser scanning microscopy and Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of EGFR and Erkl/2.
CONCLUSIONOverexpression of caveolin-1 suppresses the growth of HEp2 cells and induces apoptosis and inhibition of EGFR-MAPK signaling pathway may be involved in its mechanism.