[CCL21-CD40L fusion gene induce augmented antitumor activity in colon cancer].
Zhejiang Da Xue Xue Bao Yi Xue Ban
; 42(5): 498-503, 2013 Sep.
Article
em Zh
| MEDLINE
| ID: mdl-24167129
ABSTRACT
OBJECTIVE:
To investigate the anti-tumor activity of CCL21-exCD40L eukaryotic expression vector.METHODS:
CCL21-exCD40L fusion gene were constructed by overlap PCR connecting CCL21 and exCD40L through a flexible linker (Gly3Ser)4, and then was cloned into expression vector pcDNA3.1(+). pcDNA3.1(+)/CCL21 and pcDNA3.1(+)/exCD were constructed as negative control. Wsestern blot was used to identify the fusion protein. CHO cells was transfected with pcDNA3.1(+)/CCL21-exCD, pcDNA3.1(+)/CCL21 and pcDNA3.1(+), respectively. The chemotatic function of the expressed product was detected by Transwell method and its anti-tumor activity was tested with vivo transfection.RESULTS:
Gene sequencing and restrictive digestion proved the successful construction of pcDNA3.1(+)/CCL21-exCD40L,and its expression was conformed by western blot. The transfectant supernantes of pcDNA3.1(+)/CCL21-exCD40 group had a significant chmotactic function to DCs, of which the cell numbers passing through the film was 14.95 times of blank control every high power microscope visual field. After tumor orthotoic injection of plasmid carrying fusion gene in Balb/c mouse, the tumor mass reduced remarkablely, and all the mouse in fusion gene group survived after 4 weeks.CONCLUSION:
CCL21-exCD40L fusion protein had a remarkable function to DCs and it can inhibit tumor growth and prolong the mouse survival time, which is more effective than all control group.
Buscar no Google
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Células Dendríticas
/
Proteínas Recombinantes de Fusão
/
Neoplasias do Colo
/
Ligante de CD40
/
Quimiocina CCL21
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
Zh
Revista:
Zhejiang Da Xue Xue Bao Yi Xue Ban
Assunto da revista:
MEDICINA
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
China