| ObjectiveOral carcinoma is a common malignant tumor.It suggests drug resistance during chemotherapy,which seriously interferes with the efficacy and affects the prognosis of patients.The objective of this study was to explore the effect of salinomycin on drug resistant oral carcinoma,and to evaluate the effect of drugs on tumors from the perspectives of metastasis,invasion and drug resistance,so as to deepen the research on salinomycin.On the other hand,this study also attempts to evaluate the effect of salinomycin combined with other drugs and explore the cellular mechanism of drug action on tumors,so as to provide further theoretical guidance for its future clinical application.Method1.CCK-8 assay was used to detect the effects of salinomycin or combination of salinomycin and vincristine on proliferation of drug resistant oral epidermoid cell carcinoma(KB-V)cells;2.Trans-well was used to evaluate the effect of salinomycin on the invasiveness of KB-V cells;3.The effect of salinomycin on cell migration of KB-V cells was evaluated by scratch test;4.Flow cytometry was employed to detect the effect of salicycin and vincristine on the cell cycle of KB/V cells;5.The effects of salinomycin and glutathione on the level of reactive oxygen species in KB-V cells were evaluated by fluorescence microscope and flow cytometry;6.To determine influence of glutathione on salinomycin‘s antitumor effect;7.To evaluate the effect of salinomycin on drug resistance and ABC transporter activity in KB-V cells.Results1.CCK-8 showed that under the same concentration of vincristine,the cell survival rate of the combined salinomycin and vincristine group was significantly lower than that of the vincristine alone group;2.The scratch test showed that the cell migration rate of salinomycin group was significantly lower than that of the control group at 12-h and 24 h time point;3.Transwell experiment showed that the number of cells penetrating the polycarbonate membrane in the 4μM SAL and 8μM SAL groups were significantly lower than that of the control group.Meanwhile,the relative cell invasion rate of the cells in the 8μM SAL group was significantly lower than 4μM SAL;4.The results of flow cytometry showed that the ratio of cells in the G2/M phase of the vincristine alone group was significantly higher than that in the control group.After the combined use of vincristine and salinomycin,the ratio of cells in the G2/M phase would appear salinomycin concentration-dependent decrease;5.Fluorescence microscopy and flow cytometry results showed that salinomycin can up-regulate the level of cellular reactive oxygen species,and this up-regulation can be reversed by glutathione;6.The results of CCK-8 showed that after vincristine alone treatment,the cell survival rate dropped to 79%.When it was combined with salinomycin,the cell survival rate dropped to 36%,After the combination of vincristine,salinomycin and glycosides,the survival rate of KB/V cells rose to 77%;7.ATP-binding cassette transporter activity assay showed that the intracellular fluorescence intensity of the salinomycin group was significantly higher than that of the control group,and the cell MAF of the MRP inhibitor group was significantly higher than both P-gp and the BCRP inhibitor group.Conclusions1.Salinomycin effectively increases the sensitivity of KB/V cells to vincristine;2.Salinomycin significantly inhibits the invasion and migration ability of KB/V cells;3.Salinomycin could reverse the G2/M block of KB/V cells induced by vincristine;4.Salinomycin could reverse the drug resistance of KB/V cells by up-regulating the level of active oxygen in KB/V cells;5.Salinomycin could reverse the drug resistance of KB/V cells by inhibiting the activity of ATP-binding cassette transporter. |
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