Construction of recombinant hING4 gene and its inhibitory effect on the growth of lung cancer NCI-H460 cells
10.3781/j.issn.1000-7431.2008.11.008
- Author:
Guang-Jun DUAN
1
Author Information
1. Department of Respiration Medicine
- Publication Type:Journal Article
- Keywords:
Genes, tumor suppressor;
ING4 protein, human;
Lung neoplasms;
Mutagenesis, site-directed
- From:
Tumor
2008;28(11):942-945
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct a recombinant human inhibitor of growth 4 (hING4) gene and observe its growth inhibition effect on lung adenocarcinoma NCI H460 cells. Methods: The GFP-labeled recombinant adenovirus vector hING4 (Ad-hING4) was constructed using pcDNA3. 0-mING4 plasmid as a template and site-specific mutagenesis technique. RT-PCR and fluorescence microscope were used to detect the expression of hING4 in NCI H460 cells. Bax and Bcl-2 protein expression were detected by Western blotting. Colony formation assay and flow cytometry were used to observe the effect of hING4 on the proliferation and apoptosis of H460 cells. Results: DNA sequence analysis and PCR indicated that Ad-hING4 were successfully constructed. Ad-hING4 was expressed in lung cancer H460 cells and had obvious cytopathic effect (CPE). Western blotting suggested that hING4 gene caused up-regulation of Bax expression and down-regulation of Bcl-2 expression. Colony formation assay and flow cytometry showed that hING4 inhibited the proliferation of H460 cells and induced apoptosis of lung cancer cells. Conclusion: Ad-hING4 gene was successfully constructed. It inhibited the growth of lung cancer H460 cells in vitro.
