Impact Of XIAP Expression On HCC Chemotherapy-resistance

Objective:To investigate the effect of XIAP (X-linked inhibitor of apoptosis protein) expression on the HepG2 cells' apoptosis induced by 5-fluorouracil(5-Fu) at different concertration. Methods:The eukaryotic expression vector including full length XIAP gene was a gift from Professor David Vaux.The vector was reclaimed and transformed into the competent Escherichia coli DH5-a.The isolated vector was tested by double-enzyme cleavage method and PCR. After the vector's availability was confirmed,it was transferred into HepG2 cells by lipofectin 2000. Reverse transcription-PCR and Western blotting were used to examine the mRNA and protein expression of XIAP. Different concerntration of 5-Fu was used to detect the inhibitory rate of HepG2 cells after transfection. Result:1. Double-enzyme cleavage reaction and PCR all showed a single specific fragments,which confirmed the availability of the vector.2.Reverse transcription-PCR showed XIAP expression of transferred group was higher than the control group(p<0.05).3. Western blotting showed the XIAP protein expression was higher than the control group(p<0.05).4. HepG2 cells'inhibitory rate induced by 5-Fu at different concerntration of experimental group was all lower than control group. Conclusion:1.The expression of XIAP in HepG2 cells can be enhanced by gene engineering method.2.The XIAP could decrease the apoptosis rate of HepG2 cells induced by 5-Fu, and it could be a mechanism of chemotherapy resistant in liver cancer.