ABSTRACT
Objective: To study the effect of evodiamine (EVO) on the proliferation, cell cycle, and multidrug resistance (MDR) of leukemia K562 and K562/Adr cells. Methods: The effect on the proliferation was detected by CCK-8 kit and/or daunorubicin (DNR). The resistance index (RI) and reversal fold (RF) were calculated. The effect of EVO on cell cycle was tested by flow cytometry; Flow cytometry was used to check the fluorescence intensity of DNR. The expression of MDR1 gene in leukemia K562 and K562/Adr cells was detected by q-PCR. The expression of MDR1 and BCRP proteins in leukemia K562 and K562/Adr cells were detected by Western blotting. Results: CCK-8 test results showed that the proliferation of K562 and K562/Adr cells induced by EVO and DNR was inhibited in a dose and time dependent manner. The RI of DNR on K562/Adr cells was 30.54, the RI of EVO on K562/Adr cells was 19.09|, compared with K562 cells; The RF of DNR + EVO on K562/Adr cells was 12.07.The expression of BCRP and MDR1 protein of K562/Adr cells was significantly decreased in K562/Adr cells induced by EVO and DNR. Conclusion: Therefore EVO can effectively reverse the DNR resistance in K562/Adr cells, which may be related to reduce the expression of MDR1 on cell membrane.
ABSTRACT
Objective To investigate the reversal effect of triazole antifungal itraconazole,fluconazole combined with adriamycin on multidrug resistance in leukemia cells.Methods Human chronic myelogenous leukemia adriamycin resistant cell lines K562/ADR cells were incubated with itraconazole,fluconazole,or PSC833 (positive control) combined respectively with adriamycin.CCK-8 assay was used to assess cell proliferation of K562/ADR.The mean fluorescence intensity of intracellular ADR was measured by flow cytometry.The marker of DNA damage γH2AX was detected by Western blot.Results 1 μg/ml itraconazole and 0.5 μg/ml PSC833 can decrease K562/ADR IC50 of adriamycin from 38.30 μg/ml to 8.59 μg/ml and 24.64 μg/ml in a dose-dependent manner.K562/ADR cells were incubated with 1 μg/ml itraconazole or 0.5 μg/ml PSC833 combined respectively with adriamycin for 3 h and 6 h,the mean fluorescence intensity of intracellular ADR were increased 1.54-fold (3 h),1.50-fold (6 h) or 5.97-fold (3 h),5.83-fold (6 h).Itraconazole or PSC833 combined with adriamycin significantly increase the expression of γH2AX in K562/ ADR cells.Conclusion Itraconazole can recover adriamycin sensitivity of K562/ADR by increasing the concentration of intracelullar adriamycin and synergistically increasing DNA damage,but not for fluconazole.