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AIM:To evaluate the therapeutic efficiency of Tumor NecrosisFactor-related Apoptosis-inducing Ligand (TRAIL) gene onhuman colorectal cancer cell line HT29.METHODS:Human embryonal kidney cells transformedby introducing sheared fragments of Ad5 DNA (293 cell)were used for amplification of adenoviral vectors:Ad/GT-TRAIL,Ad/GT-Bax,Ad/GT-LacZ and Ad/PGK-GV16.Humancolorectal cancer cell line HT29 was transfected with binaryadenovirus-mediated TRAIL gene.Bax gene was used aspositive control,LacZ gene was used as the vector control,and cells treated with PBS only were used as a mockcontrol.The morphological changes,cell growth andapoptosis were measured by reversmicroscope,MTTmethod and flow cytometry.RESULTS:All adenoviral vectors titer determined by opticalabsorbency at A260nm were 1×10~(10) viral particle/ml(vp/ml).Obviously morphological changes of HT29 cells wereobserved when infected with Ad/GT-TRAIL,and thesechanges were much more obviously when Ad/PGK-GV16was coinfected.The cell suppression percentage and thepercentage of apoptotic cells were 52.5 % and 16.5 %respectively when infected with Ad/GT-TRAIL alone,whilecombining with Ad/PGK-GV16,the growth of HT29 wassuppressed by 85.2 % and the percentage of apoptotic cellswas 35.9 %.It showed a significantly enhanced therapeuticefficiency with binary system (P<0.05).CONCLUSION:A binary adenoviral vector system providesan effective approach to amplify viral vectors that expresspotentially toxic gene,TRAIL.Ad/GT-TRAIL showed asignificantly enhanced therapeutic efficiency for HT29 whencoinfected with Ad/PGK-GV16.Ad/GT-TRAIL could induceapoptosis of HT29 and inhibit its growth.
AIM: To evaluate the therapeutic efficiency of Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL) gene onhuman colorectal cancer cell line HT29. METHODS: Human embryonal kidney cells transformed by introducing sheared fragments of Ad5 DNA (293 cell) were used for amplification of adenoviral Vectors: Ad / GT-TRAIL, Ad / GT-Bax, Ad / GT-LacZ and Ad / PGK-GV16.Humancolorectal cancer cell line HT29 was transfected with binaryadenovirus- mediated TRAIL gene.Bax gene was used as positive control, LacZ gene was used as the vector control, and cells treated with PBS only were used as a mockcontrol.The morphological changes, cell growth andapoptosis were measured by reversmicroscope, MTTmethod and flow cytometry .RESULTS: All adenoviral vectors titer determined by opticalabsorbency at A260nm were 1 × 10 ~ (10) viral particle / ml (vp / ml). Obviously morphological changes of HT29 cells wereobserved when infected with Ad / GT-TRAIL, and thesechanges were much more obviously when Ad / PGK- GV16was coinfected.The cell suppr ession percentage and the percentage of apoptotic cells were 52.5% and 16.5% respectively when infected with Ad / GT-TRAIL alone, whilecombining with Ad / PGK-GV16, the growth of HT29 wassuppressed by 85.2% and the percentage of apoptotic cellswas 35.9% showed a significantly enhanced therapeuticefficiency with binary system (P <0.05). CONCLUSION: A binary adenoviral vector system provides effective approach to amplify viral vectors that expresspotentially toxic gene, TRAIL. Ad / GT-TRAIL showed asificificially enhanced therapeutic efficiency for HT29 when coinfected with Ad /PGK-GV16.Ad/GT-TRAIL could induceapoptosis of HT29 and inhibit its growth.