Mechanism of tripterine enhances the inhibitory effects of 5-fluorouracil on the proliferation and invasion in human hepatoma cells / 国际中医中药杂志

ABSTRACT

Objective To investigate the effect of tripterine combined with fluorouracil on the proliferation,invasion and apoptosis of human hepatocellular carcinoma Hep3B cells,and to identify whether the combining application of the two drugs has synergistic inhibitory effect.Methods The Hep3B cells were divided into blank control group,tripterine (2.5 mol/L) treated group,fluorouracil (25 g/ml) treated group and combined treated group (2.5 mol/L of tripterine and 25 g/ml of fluorouracil) by random number table method.Proliferation,invasion and apoptosis of different drugs treated Hep3B cells were assessed by MTS,Transwell assay,and flow cytometry,respectively.The migration differences of the four groups were detected by the scratch assay.Finally,the levels of the proliferation-related proteins p-AKT,p-ERK and the apoptosis-related proteins cleaved caspase-3 and Bax were detected by Western blotting at 24 and 48 h.Results Compared with the blank control group,the cell proliferation rate (24 h:0.305％ ± 0.016％ vs.0.768％ ± 0.063％;48 h:0.201％ ± 0.008％ vs.1.111％ ± 0.037％),and the cell migration rate (24 h:0.20％ ± 0.03％ vs.0.40％ ± 0.04％,48 h:0.25％ ± 0.02％ vs.0.59％ ± 0.07％) of combined treated group decreased (P＜0.01),while the cell apoptosis rate (24 h:24.33％ ± 3.85％ vs.3.80％ ± 0.40％,48 h:45.10％ ± 4.10％ vs.8.47％ ± 1.65％) significantly increased (P＜0.01).Compared with the blank control group,the cleaved caspase-3 (24 h:1.39 ± 0.11 vs.1.01 ± 0.04,48 h:1.38 ± 0.12 vs.0.99 + 0.03) and Bax (24 h:1.35 ± 0.13 vs.1.00 ± 0.08,48 h:1.39 ± 0.09 vs.0.99 ± 0.05) protein expression of combined treated group significantly increased (P＜0.05).After 24 h of the intervention,the number of cell invasionin (15 ± 6 vs.231 ± 38) the combined group was significantly lower than that in the blank control group (P＜0.05).The protein expression of p-AKT/AKT (0.79 ± 0.04 vs.0.99 ± 0.05) and p-ERK/ERK (0.52 ± 0.04 vs.0.75 ± 0.07) in the combined group was significantly lower than that in the fluorouracil group 48 h after the intervention (P＜0.05).Conclusions The combination application of 5-FU and Tripterine on Hep3B cells significantly inhibited the cells proliferation,migration and invasion ability and promoted the apoptosis of Hep3B cells by down-regulating the expression of p-AKT and p-ERK and up-regulating the expression of cleaved caspase-3 and Bax.