Abstract
BackgroundThe development of new modulator possessing high efficacy, low toxicity and high selectivity is a pivotal approach to overcome P-glycoprotein (P-gp) mediated multidrug resistance (MDR) in cancer treatment. In this study, we suggest a new molecular mechanism that TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) down-regulates P-glycoprotein (P-gp) through inhibition of DNA-PKcs/Akt/GSK-3β pathway and activation of caspases and thereby sensitize MDR cells to MDR-related drugs.ResultsMDR variants, CEM/VLB10-2, CEM/VLB55-8 and CEM/VLB100 cells, with gradually increased levels of P-gp derived from human lymphoblastic leukemia CEM cells, were gradually more susceptible to TRAIL-induced apoptosis and cytotoxicity than parental CEM cells. The P-gp level of MDR variants was positively correlated with the levels of DNA-PKcs, pAkt, pGSK-3β and c-Myc as well as DR5 and negatively correlated with the level of c-FLIPs. Hypersensitivity of CEM/VLB100 cells to TRAIL was accompanied by the activation of mitochondrial apoptotic pathway as well as the activation of initiator caspases. In addition, TRAIL-induced down-regulation of DNA-PKcs/Akt/GSK-3β pathway and c-FLIP and up-regulation of cell surface expression of death receptors were associated with the increased susceptibility to TRAIL of MDR cells. Moreover, TRAIL inhibited P-gp efflux function via caspase-3-dependent degradation of P-gp as well as DNA-PKcs and subsequently sensitized MDR cells to MDR-related drugs such as vinblastine and doxorubicin. We also found that suppression of DNA-PKcs by siRNA enhanced the susceptibility of MDR cells to vincristine as well as TRAIL via down-regulation of c-FLIP and P-gp expression and up-regulation of DR5.ConclusionThis study showed for the first time that the MDR variant of CEM cells was hypersensitive to TRAIL due to up-regulation of DR5 and concomitant down-regulation of c-FLIP, and degradation of P-gp and DNA-PKcs by activation of caspase-3 might be important determinants of TRAIL-induced sensitization of MDR cells to MDR-related drugs. Therefore, combination of TRAIL and chemotherapeutic drugs may be a good strategy for treatment of cancer with multidrug resistance.
Highlights
The development of new modulator possessing high efficacy, low toxicity and high selectivity is a pivotal approach to overcome P-glycoprotein (P-gp) mediated multidrug resistance (MDR) in cancer treatment
The high susceptibility of MDR cells to TNF-related apoptosis inducing ligand (TRAIL) is associated with up-regulation of DR5 and down-regulation of Cellular FLICE inhibitory protein (c-FLIP) in the cells Many studies have focused on overcoming MDR through modulation of P-gp function [4,26]
In a model of CEM cells with gradually increasing expression of multidrug-resistance 1 (MDR1) gene, the higher expression level of MDR1 gene in MDR variants, the higher cytotoxicity to TRAIL as shown in MTT assay (Figure 1A) and flow cytometric analysis of apoptosis (Figure 1B), demonstrating that the MDR cells were more susceptible to TRAIL-induced cell death than TRAIL-insensitive parental cells
Summary
The development of new modulator possessing high efficacy, low toxicity and high selectivity is a pivotal approach to overcome P-glycoprotein (P-gp) mediated multidrug resistance (MDR) in cancer treatment. We suggest a new molecular mechanism that TRAIL (tumor necrosis factor-related apoptosis-inducing ligand) down-regulates P-glycoprotein (P-gp) through inhibition of DNA-PKcs/Akt/GSK-3b pathway and activation of caspases and thereby sensitize MDR cells to MDR-related drugs. Inhibition of P-gp function or expression can reverse P-gp-mediated MDR and improve the efficacy of chemotherapy [9]. It has been demonstrated that DNA-PKcs-catalyzed RNA Helicase A phosphorylation enhanced the transcription of the MDR1 gene through the CAAT-like element of the MDR1 gene promoter and DNA-PKcs played an important role in regulation of P-gp expression by MDR1 promoter activation [12]
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