| ObjectiveLiver cancer systemic treatment although many, but the effect is not optimistic,the survival rate is Still low, the metformin, oxaliplatin, and the combination of the two arerespectively applied to BEL-7402cells, and set the corresponding observed in controlgroup, the observation of drug different effects on the cells, and observe whether the drughas a synergistic effect, so as to provide basis for Clinical treatment.MethodsTo culture BEL-7402cells in vitro and divided into four groups:control,metformin-treated,oxaliplatin-treated and combination group. Chosing the growing cellsfor experiment,The cell growth inhibition rate after treated with the drug (MTT method).Using the electron microscopy to observe the changes in cell morphology andultrastructure in the role of the drugs. Observation of the cell cycle and cell apoptosis bythe flow cytometry. Using Western Blot Technology to observe caspase-3ã€caspase-9andthe expression of the pro apoptotic protein Bax expression and anti apoptotic protein Bcl-2.Results1.Different concentrations of metformin, oxaliplatin alone and in combination caninhibit the growth of BEL-7402cells. In a separate case of drugs, increase over time, theinhibiting effect on cell more obviously, there was significant difference (P<0.05); thecombination of the two drugs, compared with the single drug, the inhibition rate increasedsignificantly, and the inhibition rate were significantly different (P<0.05).2.Electron microscopy revealed that metformin and oxaliplatin on BEL-7402cell, Thecell adherence ability decline, decrease in cell size, roundness, condensation of cytoplasm,nuclear pyknosis, cell floating in the culture fluid, showed morphological changes at theearly stage of apoptosis. 3. Flow cytometry results show: the48h after drug treatment, flow cytometry showed,metformin group and oxaliplatin group of apoptosis rate were17.63±1.12%and20.13±0.86;compared with the blank control group (13.15±0.65) had significant difference (P<0.05),apoptosis of the combination group was27.42±1.05%; compared to with the drug alone,there was statistically significant difference (P<0.05).4.Detection of Western Blot technology, the expression and activity of Caspase-3,caspase-9were significantly increased in the metformin and oxaliplatin effect, in thecombination of the two case of drugs, the effect is more obvious. In the role of the drugapoptosis protein Bax expression was significantly increased, while the expression of antiapoptotic protein Bcl-2in single and combination cases showed no significant changes.Conclusions1.Metformin and oxaliplatin alone or combined application can inhibit theproliferation of human hepatoma cell line BEL-7402, promote the apoptosis.2.Compared with the single drug,the combined effect of metformin and oxaliplatin aremore obvious.With the increase of time, the effect is more obvious.3.Metformin and oxaliplatin for apoptosis of hepatoma cells by Caspase cascade, andmay be related to the increase of the pro apoptotic protein Bax. |
