Inhibition Effect of Ad-IL24 Combined with Low Dose of 5-Fu on Hepatocarcinoma Cells HepG2 and Its Mechanism

Objective To investigate the inhibition effect of Ad-IL24 combined with low dose of 5-Fu on hepatocarcinoma cells HepG2 and its mechanism.

Methods PCR, restriction enzyme cleavage and linkage were used to construct the adenovirus expression vector pAd-IL24. The expression of IL24 was detected by Western blot. CCK8 assay was applied to confirm the doses of 5-Fu and to detect the viability of HepG2 cells which were treated by Ad-IL24 with or without 5-Fu. The proportion of GFP positive cells in HepG2 cells treated by different does of 5-Fu and adenovirus was detected by flow cytometry. The expression levels of coxsackie adenovirus receptor (CAR) were detected by flow cytometry before and after 5-Fu treatment.

Results The adenovirus expression vector pAd-IL24 was successfully constructed. The doses of 5-Fu were confirmed to be 1 and 2μg/ml by CCK8 assay; Ad-IL24 combined with low doses of 5-Fu synergistically inhibited the growth of HepG2 cells, and the combination index were 0.75 and 0.64 respectively. After treated with low dose of 5-Fu, the level of CAR expression on HepG2 cells was significantly increased(P<0.01).

Conclusion The combination treatment of Ad-IL24 and low dose of 5-Fu synergistically inhibit the growth of hepatocarcinoma cells HepG2, which maybe related with up-regulation of CAR induced by 5-Fu.