Human Id3 expression in lung adenocarcinoma cell line A549 and its inhibiory effect on cell growth
- VernacularTitle:人Id3基因在肺腺癌A549细胞中的表达及其对细胞生长的抑制
- Author:
Chuandong ZHU
- Publication Type:Journal Article
- Keywords:
Inhibitor of differentiation 3(Id3)gene;
lung neoplasms cell;
proliferation repression;
apoptosis
- From:
Chinese Journal of Cancer Biotherapy
2006;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the inhibitory effect of inhibitor of differentiation 3(Id3)on growth of human lung adenocarcinoma cell line A549.Methods:Recombinant eukaryotic expression vector pEGFP/Id3 was constructed and transfected into A549 cells by liposome-mediated method.Expression of pEGFP/Id3 in A549 cells was analyzed by flow cytometry(FCM),fluorescence microscopy,semi-quantitative RT-PCR and immunocytochemistry.The growth inhibitory rate of A549 cells was examined by MTT assay;cell cycle change was evaluated by PI(propidium iodide)staining method.Cell apoptotic rate and nuclear morphology were detected by Annexin V/7-AAD and Hoechst33258 staining. Results:The recombinant eukaryotic expression vector pEGFP/Id3 was successfully constructed.The expression of EGFP reached the peak 48-72 h after transfection;the expresion of pEGFP-transfected group was higher than that of the pEGFP/ Id3 group.RT-PCR and immunocytochemistry staining showed that Id3 mRNA and protein were effectively expressed in pEGFP/Id3-transfected A549 cells.The growth of cells in pEGFP/Id3 transfeeted cells was significantly inhibited 48-72 h after transfection(P
