Construction of recombinant adenovirus containing p53 up-regulated modulator of apoptosis and its effect on pancreatic cancer in vitro and in vivo
10.3760/cma.j.issn.0254-5101.2009.01.016
- VernacularTitle:p53正向凋亡调节因子重组腺病毒载体的构建及对胰腺癌细胞增殖的抑制作用
- Author:
Kejun ZHANG
;
Dechun LI
;
Haining CUI
- Publication Type:Journal Article
- Keywords:
Adenovirus vector;
p53 up-regulated modulator of apoptosis(PUMA);
Gene therapy;
Apoptosis;
Pancreatic carcinoma
- From:
Chinese Journal of Microbiology and Immunology
2009;29(1):65-70
- CountryChina
- Language:Chinese
-
Abstract:
objective To construct the recombinant adenovirus containing p53 up-regulated modulator of apoptosis(Ad-PUMA)and investigate its growth inhibition effect on pancreatic callCer cells in vitro and in vivo.Methodls Ad-Easy system was used to construct Ad-PUMA by recombination in E.coli.The virus was Dackaged in 293 cells and subsequently identified valid.The AsPC-1 cells were infected with AdPUMA.Before and after Ad-PUMA infection,the expression of PUMA protein wag investigated by western blot,the inhibition rate of AsPC-1 cells was examined by MTY assay.The in vivo tumor suppressive effect was detected in nude mice with human AsPC-1 xenograft.PUMA protein and the apoptosis of AsPC-1 xenograft were detected by western blot and TUNEL(terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling)method.Results In vitro,the expression of PUMA protein was increased with titer of Ad-PUMA,the proliferation of AsPC.1 cells were suppressed,significantly,and the effect was in a viral dose-dependent nlanner.In vivo,the growth in nude mice of AsPC-1 infected with Ad-PUMA was significantly inhibited with an inhibition rate of 44.2%.The expression of PUMA was significantly up-regulated,and the apoptosis index wa8 significantiv increased in tumor after Ad-PUMA infection as determined by western blot and TUNEL.Conclusion The expression of PUMA call inhibit the proliferation of pancreatic cancer in vitro and in vivo,and may be used 88 a potential tool for cancer therapy
