Study on the effect of costunolide on sensitivity of chronic myeloid leukemia K562/ADR cells to doxorubicin via p38-MAPK pathway / 白血病·淋巴瘤

ABSTRACT

Objective:To explore the effect of costunolide on sensitivity of chronic myeloid leukemia cell line K562/ADR to doxorubicin and its mechanism.Methods:K562/ADR cells in the logarithmic phase were used, and the cells were treated with different concentrations of costunolide, doxorubicin or costunolide combined with doxorubicin for 72 h. The cell proliferation was detected by CCK-8 method, the cell proliferation rate was calculated, and the half inhibitory concentration (IC 50) of the two drugs was obtained. The cells were treated with 10 μmol/L costunolide, 10 μmol/L doxorubicin or costunolide combined with doxorubicin for 48 h, the apoptotic rate was detected by flow cytometry, and the expression level of p38-MAPK pathway related proteins was detected by Western blot. Results:The cell proliferation rate in the costunolide combined with doxorubicin group was lower than that in the corresponding concentration of the two drugs alone groups, and the differences were statistically significant both ( P < 0.05). The IC 50 of doxorubicin for K562/ADR cells was (13.50±0.86) μmol/L, and costunolide was (7.30±0.55) μmol/L ( t = 7.044, P = 0.002). The results of flow cytometry showed that the apoptosis rate of K562/ADR cells in the 10 μmol/L costunolide combined with 10 μmol/L doxorubicin group was higher than that of the blank control group, costunolide alone group and doxorubicin alone group, and the differences were statistically significant [(19.68±3.21)% vs. (2.96±0.87)%, (9.34±2.89)%, (9.18±2.13)%, all P<0.01]. Compared with the 10 μmol/L costunolide alone group and the 10 μmol/L doxorubicin alone group, the expression of the apoptosis inhibitor protein bcl-2 in the two-drug combination group was down-regulated, and the expressions of bad, p-p38, cleaved-caspase-3 and cleaved-PARP proteins were up-regulated. Conclusion:Costunolide can enhance the inhibitory effect of doxorubicin on the proliferation of K562/ADR cells and promote doxorubicin-induced apoptosis, which may reverse the drug resistance of K562/ADR cells by regulating the p38-MAPK pathway.