title: Adenovirus mediated BIMS transfer induces growth supression and apoptosis in Raji lymphoma cells.
abstract: OBJECTIVE
  To transfer pro-apoptotic BIM directly into tumor cells bypass the complicated biological processes of BIM activation so as to reverse the chemoresistance of cancer cells.
  
  
  METHODS
  BIMS was specifically amplified from HL-60 cells by RT-PCR, confirmed to be correct by sequencing and cloned into shuttle vector pAdTrack-CMV carrying a green fluorescence protein gene to generate a recombinant plasmid pAdTrack-CMV-BIMS. This plasmid and adenovirus backbone plasmid pAdEasy-1 were linearized and electroporated into E.coli BJ5183 host bacteria to mediate homologous recombination. The positive clone was identified by restrict endonuclease digestion. The recombinant pAdEasy-CMV-BIMS was transferred into HEK293 cells for packaging and amplification. The successful construction of recombinant human BIMS adenovirus (Ad-BIMS) was demonstrated by Western blot. To test whether Ad-BIMS has the capability of inducing apoptosis of tumor cells, Ad-BIMS was used to infect GC resistant Burkitt lymphoma Raji cells.
  
  
  RESULTS
  After infected for 2-5 days, BIMS expression in Raji cells was detected by RT-PCR and Western blot. The significant growth retardation and apoptosis of Raji cells were also observed by MTT and flow cytometry.
  
  
  CONCLUSIONS
  These results indicated that BIMS might be a potential candidate of gene therapy for chemoresistant tumor cells.
authors.affiliation: Department of Pediatric Hematology and Immunology, West China Second University Hospital, Sichuan University. Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, Chengdu 610041, Sichuan, China; Department of Phymatology, Baoji Municipal Central Hospital, Baoji 721008, Shaanxi, China.
authors.forename: Ya Ning
authors.initials: YN
authors.lastname: Zhao
authors.affiliation: Department of Pediatric Hematology and Immunology, West China Second University Hospital, Sichuan University. Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, Chengdu 610041, Sichuan, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
mesh_terms: D000256:Adenoviridae; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000235:genetics*; D000072224:Bcl-2-Like Protein 11; D002051:Burkitt Lymphoma / Q000628:therapy*; D015316:Genetic Therapy; D005822:Genetic Vectors; D057809:HEK293 Cells; D018922:HL-60 Cells; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics*
chemical_list: D051017:Apoptosis Regulatory Proteins; C000606775:BCL2L11 protein, human; D000072224:Bcl-2-Like Protein 11; D008565:Membrane Proteins; D011518:Proto-Oncogene Proteins
keywords: Apoptosis; BIMS; Chemoresistance; Gene therapy; Lymphoma; Recombinant adenovirus; Transfection; Tumor