<p><b>OBJECTIVE</b>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.</p><p><b>
METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>These results indicated that BIMS might be a potential candidate of
gene therapy for chemoresistant
tumor cells.</p>