[Gene therapy for melanoma by bifidobacterium infantis-mediated transfer of CD and UPRT genes with 5-FC in vitro].
OBJECTIVE
This study was conducted by using Bifidobacterium Infantis as a delivery system to transport suicide gene CD and UPRT to tumors in an attempt to assess the UPRT-enhanced antitumor effect of CD/5-FC.


METHODS
The recombinant plasmid pGEX-UPRT was constructed and transfered into Bifidobacterium Infantis by electroporation and then identified. The synergistic antitumor effect of coexpression CD and UPRT was determined by MTT method. And the morphologic changes of B16-F10 cells were observed.


RESULTS
Recombinant Bifidobacterium Infantis could express UPRT correctly. The suvival rate of cells administrated CD+ UPRT and 5-FC was significantly lower than that of control (P<0.01), and the 5-FC sensitivity (IC50 = 0.015 micromol/mL) exhibited a 8. 5-fold increase when compared with that of cells administrated CD alone (IC50 = 0.127 micromol/mL). The cells treated with CD+UPRT were remarkably damaged morphologically, and the growth of cells was significantly inhibited as compared with that of other groups.


CONCLUSIONS
Recombinant Bifidobacterium Infantis with UPRT gene can significantly enhance the killing effect of CD/5-FC suicide gene system on melanoma B16-F10 cells of mice.
An
Li-na
LN
Pathophysiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Li
Zhu-hua
ZH
Yue
Yang
Y
Wang
Shu-ren
SR
Guo
Zhi-ying
ZY
D000818:Animals; D001644:Bifidobacterium / Q000235:genetics*; D045744:Cell Line, Tumor; D017077:Culture Media, Conditioned / Q000494:pharmacology; D043525:Cytosine Deaminase / Q000235:genetics*; D005437:Flucytosine / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D041722:Genes, Transgenic, Suicide / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D008545:Melanoma / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D051379:Mice; D010430:Pentosyltransferases / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping
D017077:Culture Media, Conditioned; D012333:RNA, Messenger; D005437:Flucytosine; D010430:Pentosyltransferases; C023269:uracil phosphoribosyltransferase; D043525:Cytosine Deaminase