Construction of siBcl2-Ercc1 recombinant adenovirus and its effects on apoptosis of lung cancer cells
10.3969/j.issn.1006-5725.2015.15.010
- VernacularTitle:重组人Bcl2及Ercc1基因腺病毒的构建及对肺癌细胞凋亡的影响
- Author:
Fang WANG
;
Jin LIU
- Publication Type:Journal Article
- Keywords:
Lung cancer;
Apoptosis;
Recombinant adenovirus;
H460
- From:
The Journal of Practical Medicine
2015;31(15):2443-2446
- CountryChina
- Language:Chinese
-
Abstract:
Objective Toexplore the effect of Bcl2 and Ercc1 gene on apoptosis by RNA interference of Bcl2 and Ercc1 gene expression in non-small cell lung cancer cell line. Methods shRNA targeting Bcl2 and Ercc1 gene were cloned into vector to construct recombinant adenovirus plasmids which were transfected into HEK293 cells. High titer adenovirus was obtained after package and amplification. H460 cells were infected with the adenovirus and apoptosis rate was detected by flow cytometry. Results The results of restriction endonuclease digestion showed that recombinantadenovirus vector adsiBcl2-Ercc1 was successfully constructed. The positive 1 200 bp amplification bands could be seen in PCR analysis , and the titer of the adenovirus was 5 × 1011/mL. The apoptosis rate of AdsiBcl2-Ercc1 was higher than that of the two single silenced gene groups. Conclusions The recombinant plasmids and recombinant adenovirus were successfully constructed. Silencing of Bcl2 and Ercc1 through shRNA promotes the apoptosisin lung cancer cells.
