title: [CCL21-CD40L fusion gene induce augmented antitumor activity in colon cancer].
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Oncology, Tianjin General Hospital, Tianjin 300052, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Gong
authors.forename: Hong-Li
authors.initials: HL
authors.lastname: Zhou
authors.forename: Yi
authors.initials: Y
authors.lastname: Ba
mesh_terms: D000818:Animals; D023201:CD40 Ligand / Q000235:genetics / Q000494:pharmacology*; D016466:CHO Cells; D045744:Cell Line, Tumor; D054421:Chemokine CCL21 / Q000235:genetics / Q000494:pharmacology*; D003110:Colonic Neoplasms / Q000628:therapy*; D003412:Cricetulus; D003713:Dendritic Cells / Q000187:drug effects* / Q000502:physiology; D015316:Genetic Therapy; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*
chemical_list: C515478:CCL21 protein, human; D054421:Chemokine CCL21; D011993:Recombinant Fusion Proteins; D023201:CD40 Ligand