| Background and objective:1,2,3-Triazoles has occupied an important role not only in organic chemistrybut also in medicinal chemistry due to their easy synthesis and attractive features aswell as numerous biological activities. Recent researches demonstrate the majority ofsynthesized1,2,3-triazole groups exhibit potential activities against the growth ofMGC-803, MCF-7, PC-3and EC-109cells, which lead a growing attention toresearch them as anticancer agents. DYC-279is a newly synthesized carboxylate witha1,2,3-triazole unit and its relative molecular is468. To our best knowledge, littleresearch has been devoted to the mechanism of its anticancer effect. In this paper, weherein reported the anticancer activity and the induction effect of DYC-279on cellcycle arrest and cell apoptosis in vitro and its inhibition effect on activity of CYP450enzymes.Methods:Human hepatocellular carcinoma HepG-2cells were exposed to DYC-279of0.39,0.78,1.56,3.13,6.25,12.5mg/L for24,48or72h, CCK-8assay was used toevaluate the antitumor activities of DYC-279on HepG-2cells in vitro. After exposureto0.78,1.56,3.13mg/L of DYC-279for24h, the changes of cell cycle and apoptosiswere detected with FCM and the mRNA or protein expression levels of cyclinB1,cdc-2, cyclinD1caspase9, Bax and Bcl-2were evaluated by RT-PCR or western blot.The model of S9of rat was used to detect the effect of DYC-279on metabolicenzymes activity of CYP1A2, CYP2D6, CYP2E1in vitro.Results:(1) The antitumor activities of DYC-279in vitroThe results of CCK-8showed that as the increasing of concentration andexposed time of DYC-279, the inhibition ratio gradually increased. In the treatmentswith DYC-279, the IC50at24h was (10.6±2.7) mg/L, while the IC50was obtained ata concentration of (6.5±1.9) mg/L at48h, and (3.1±1.1) mg/L at72h.(2) The interference effect of DYC-279on cell cycle or apoptosis of HepG-2cells FCM assay results demonstrated that following treatments with differentconcentration of DYC-279for24h, there was a decrease of cells in the G0/G1phasewhile both cells in the S and G2/M increased as the dose increasing compared tocontrol; Annexin V-FITC/PI double staining results revealed the percentage of cells inearly apoptosis increased in a concentration-dependent manner while even the lowestconcentration tested caused significant apoptosis compared to control group.(3) The effect of DYC-279on cell cycle related genesHepG-2cells were exposed to different concentration of DYC-279for24h, themRNA expression levels of cyclinB1, cdc-2were up-regulated while cyclinD1wassubstantially down-regulated with increasing drug concentration.(4) The study of DYC-279on proteins related to cell cycle or apoptosis expressionWestern blot results found that cyclinB1cdc-2, caspase-9and Bax wereup-regulated while down-regulated for cyclinD1and Bcl-2after dealt with DYC-279of0.78,1.56,3.13mg/L, respectively.(5) The research of DYC-279on metabolic enzymes of CYP450in vitroCompared to control group, the non-significant change of Phenacetinconcentration demonstrated DYC-279had little effect on the activity of CYP1A2enzyme; DYC-279decreased the activity of CYP2D6by lowering the metabolism ofDextromethorphan; Similarly, the metabolism of Chlorzoxazone decreased as theincreasing of DYC-279concentration compared to control group which predicted theinfluence of DYC-279on the activity of Chlorzoxazone enzyme maybe involved in adose-depend manner.Conclusion:(1) DYC-279inhibited the proliferation of HepG-2cells.(2) DYC-279induced cell cycle arrest in G2/M phase and apoptosis.(3) The antitumor effect of DYC-279may be related to the irregularityexpression of cyclinB1, cdc2, cyclinD1, caspase9, caspase3, Bax and Bcl-2.(4) DYC-279inhibited the metabolizing enzymes activities of CYP2D6,CYP2E1but had little effect on CYP1A2. |
PDF Full Text Request