The Preliminary Study Of Effect Of Combination Of Dihydroartemisinin And Doxorubicin On Multidrug Resistance Reversal On HCT8/ADR Colon-cancer Cells

ObjectiveMultidrug resistance is barrier to the treatmen and prognosis of cancer. To investigate the effect of combination use of Dihydroartemisinin (DHA) and doxorubicin (DOX) on reversal of drug-resistance, the cell proliferation, cell cycle regulation, apoptosis, and autophagy were investigated by using the drug-resistant human colon tumor HCT8/ADR cell line.MethodsHCT8/ADR colon cancer cells were cultured in vitro.48hous later, HCT8/ADR colon cancer cells were collected and planted on plate at certain density. ①cancer cells were planted on the 96-wells plate at the density of 5X 103 cells per well.4 groups including Blank group, DOX group, DHA group and DHA+DOX group were established, setting drug concentration as follow:DOX(mg · L-1): 0、1、2、3、4、5、6、7、8、9、10; DHA(mg·L-1):0、1、2、3、4、5、6、7、8、 9、10; Combined group:DHA(mg · L-1):5, 10mg/L; DOX(mg · L-1):0.05、0.1、0.2、 0.3、0.4、0.5、0.6、0.7、0.8、0.9、1.0、1.1、1.2、1.3、1.4、1.5. After cells were cultured in 96-wells plate 24hours, MTT test was conducted to examine the inhibition effect of DHA, DOX and combination of DHA and DOX on the HCT8/T cells, respectively;②cancer cells were planted on the 12-wells plate at the density of 5×104 cells per well.4 groups including Blank group, DOX group:0.05、0.01 mg/L; DHA group:10 mg/L and DHA+DOX group:DOX group: 0.05、0.01 mg/L; DHA group:10 mg/L. Their effect on cycle regulation and apoptosis was investigated using flow cytometry, and autophagy was analyzed using Western blottingResult1. Compared with single drug use, MTT results proved that the combination of DHA (10 mg/L)and DOX(0.1 mg/L) could effectively inhibit the cell viability of HCT8/ADR below 50%.2. Compared with single drug use, Flow cytometry proved that the combination of DHA and DOX could effectively induce apoptosis of HCT8/ADR cells.3. Western blot test show that the combination of DHA and DOX can induce autophagy of HCT8/ADR cells through up-regulate the expression of LC3-Ⅱ protein and down-regulate the expression of Cyclin DlproteinConclusionThe combination of DHA and DOX could inhibit the growth of HCT8/ADR cells and promote the occurrence of apoptosis. Meanwhile, the combination of two drug could up-regulate the expression of LC3-Ⅱprotein which lead to the production of autophagosome lowering the expression Cyclin D1 protein. It suggested that DHA could reverse the multidrug resistance of HCT8/ADR and restore the cancer cell’s sensitivity to the chemotherapeutics...