Effect Of Adenovirus - Mediated ShRNA On The High Expression Of CFLIP In Colorectal Cancer Cell Line TRAIL

Objective By using recombinant adenoviruses carrying cFLIP shRNA transfection colorectal cancer cells, to investigate the effect of down-regulated cFLIP expression on colorectal cancer cells’ pesticide resistance and apoptosis.Methods 1. Chosing colorectal cancer cells SW620 to cultivate and passage.2.Using recombinant adenoviruses carrying cFLIP shRNA which regulated by hU6 promoter, transfect colorectal cancer cells by recombinant adenoviruses, and using meaningless sequence-adenoviruses as contrast.3.Use following methods to test cell data:a) Realtime-PCR were used to detect the expression of cFLIP mRNA level and Westernblot were used to detect the expression of cFLIP gene on protein level; b) Flow cytometry were used to evaluate cell apoptosis.Results After recombinant adenoviruses transfection, the expression of cFLIP mRNA level have significant difference (P<0.05) among the target gene group (0.12±0.02), the negative control group (0.38±0.01) and the blank control group (0.41±0.03); and cFLIPL protein level also have significant difference (P<0.05) among the target gene group (0.75±0.12), the negative control group (1.09±0.06) and the blank control group (1.12±0.10).And the rate of cell apoptosis have significant difference (P<0.05) among the non-infectied group (5.07±1.16%), the non-infectied+TRAIL group (12.84±2.33%), the infectied group (6.51±1.56%) and the infectied+TRAIL group (34.16±2.43%)Conclusion Colorectal cancer cells’TRAIL resistance were related to the high expression of cFLIP level, and the sensitivity to TRAIL could increased by down-regulated cFLIP expression on colorectal cancer cells.