| Objective :(1) To observe the anti-tumor effects of salinomycin combination sorafenib on three human malignant cells in vitro and to analysis the anti-cancer efficacy.(2) Liver xenograft tumor model was used to investigate the inhibition effects of salinomycin combination sorafenib, For the development of application of salinomycin treat human malignant tumors to provide experimental evidence.Methods :(1) MTT assay was applied to assess the anti-proliferative effects. The experiment is divided into six groups: blank control group,the salinomycin group, the sorafenib group, salinomycin/sorafenib 2:1group, salinomycin/sorafenib1:1 group, salinomycin/sorafenib 1:2 groups;A single drug group setting 96, 48, 24, 12, 6, 3, 1.5, 0.75, 0.375,0.1875ug/ml in total ten concentration, combination group set up 48, 24,12, 6, 3, 1.5, 0.75, 0.375, 0.1875, 0.09375ug/ml in total ten concentration combined according to the ratio. Measuring absorbance(OD Value) with a microplate reader,Using excel and ORIGIN 8.0 software to deal with data,fitting the concentration-response curves and functional equation,Loewe isobologram combination index formula CI=(dA)/(DA)+(dB)/(DB) was applied to calculate the CI value and evaluate the interactively effects of the two drugs.(2)Drug treating liver transplantation in situ tumor model;With CMV package and infecting Hep G-2 cell,establishing stable transfection cell Hep G-2-GFP-Luc, in vitro amplification, preparation into single cell suspension, injected hypodermically into nude mouse.Selected 18 to 22 g female BALB/c- nu mice, Tumor block transplanting method was used to establishing the xenograft tumors model;Spectrum CT measurement of luminous intensity value, and completely randomly divided into 4 groups: Blank control group, salinomycin 10 mg/kg group,Sorafenib 10 mg/kg group, salinomycin+sorafenib 10 mg/kg+10 mg/kg group. The way of drug delivery for the experimental group were intragastric administration.Mice were euthanized at end of the treatment,get the liver and tumor, weighing and take photos, according to the results of the tumor weight, judging the combination effect.Results: The efficacy of window was covered by the designed concentrations range.Two drugs used alone and different fixed ratio share dose-response curves were "S" type;By computing the CI value of two drugs used in combination with a fixed proportion of different cell inhibition rate is 30%, 50%, 70% and 90%, human kidney cell KCC853:sal /sor 1:1 shows additive effect or close to this effect, but with the other two proportional show antagonism effect. two drug combined on human liver cancer cell Hep G-2, sal /sor 1:2 shows additive effect or close to this effect, but with the other two proportional shows antagonism effect more.lung adenocarcinoma cells A549:all CI value are within the range of 0.9~ 1.1, belongs to the additive effect.(2)According to the observation of the Hep G-2 xenograft tumor,the data showed that the inhibition ratio of salinomycin?sorafenib and combination group is 44.5% ?48.3% and69.1%.Conclusion:(1)The proliferation inhibition strength of salinomycin and sorafenib on three kinds of tumor cells are similar; Drugs used alone or combined can strong inhibitory cancer cells.(2)The combination effect produced by salinomycin combined with sorafenib in vitro can be addition and synergism, is related to the concentration, the proportion and the cell species;(3)Salinomycin and sorafenib can significantly inhibited tumor growth in nude mice, the combination has a stronger inhibitory effect. |
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