Experimental Study on the Mechanism of Reversal of Leukemia Multidrug Resistance by Proteasome Inhibitor Bortezomib

Abstract

OBJECTIVE In this study, we applied multidrug resistant leukemia cell line expressing mdr1-mRNA to observe changes in mdr1-mRNA, the P-gp, cell cycle and apoptosis before and after bortezomib was used, in order to explore the mechanism of reversal of leukemia multidrug resistance by the proteasome inhibitor bortezomib.METHODS Flow cytometry (FCM) was used to detect the intracellular drug concentration, expression of P-gp, cell apoptosis and cell cycle status of K562/DNR cells before and afer treatment with different concentrations of bortezomib. Fluorescence quantitative PCR was applied to detect the mdr1-mRNA expression in K562/DNR and K562/S cells. RESULTS Bortezomib could increase the intracellular DNR content in K562/DNR cells, but showed no effect in K562/S cells. 5-100 nmol/L bortezomib could significantly reduce the P-gp/mdr1-mRNA expression in K562/DNR cells in vitro, and showed a dose-dependent effect. There was a statistically significant difference (P < 0.05) between different concentration groups and the control group. P-gp/mdr1-mRNA expression was negatively correlated with cell apoptosis (r = -0.912 and P < 0.01). After treatment with different concentrations of bortezomib for 24 h, K562/DNR cells in G2 + M phases were significantly increased, while cells in G0 + G1 phases and S phase were significantlydecreased, accompanied by an increased apoptotic rate.CONCLUSION Bortezomib can induce G0 + G1 phase to G2 + M phase, and thereby enhance the chemosensitivity of leukemia, and may also reverse the multidrug resistance in leukemia mediated by P-gp overexpression encoded by mdr1 gene. This confirms that bortezomib can reverse leukemia multidrug resistance at the levels of nucleic acid and protein molecules.

Keywords

• proteasome inhibitor
• bortezomib
• multi-drug