Targeting P-Glycoprotein: Nelfinavir Reverses Adriamycin Resistance in K562/ADR Cells

Abstract

Background/Aims: The emergence of multidrug resistance (MDR) caused by P-glycoprotein (P-gp) overexpression is a serious obstacle to the treatment of chronic myelocytic leukemia. In recent years, some clinical trials have shown that nelfinavir (NFV), a traditional anti-HIV drug, has anti-cancer effects. Some researchers have also shown NFV might be a potential P-gp inhibitor. This study is aimed at investigating whether nelfinavir can act as an MDR-reversal drug and to clarify its molecular mechanism as well. Methods: K562 and K562/ADR cell lines were applied in the study. Cytotoxicity was detected by CCK-8 reagents. Cell apoptosis was detected by flow cytometry and inverted fluorescence microscopy to detect the binding of apoptotic dyes to cells. Western blot was used to detect the expression of proteins. Drug-protein molecular docking simulation by using Sybyl-x 2.0 software. Results: Non-toxic concentrations of NFV (1.25–5 μM) could reverse Adriamycin (ADR), colchicine, paclitaxel, and imatinib resistance of K562/ADR cells, with reversal indexes of up to 10.8, 7.4, 57, and 9.3, respectively. NFV inhibited P-gp efflux function, as evidenced by the significant increase in the intracellular accumulation of ADR and Rho-123, without affecting P-gp protein and mRNA expression levels. Further ATP content detection and molecular docking simulations showed that NFV could decrease intracellular ATP content and has a high affinity with the active functional regions of P-gp, respectively. When co-administered with ADR, NFV increased intracellular reactive oxygen species as well as blocked the ERK/Akt signaling pathway, leading to cell apoptosis. Conclusion: NFV inhibited P-gp function, decreased intracellular ATP content, and promoted cell apoptosis in K562/ADR cells, thereby reversing MDR. These findings encourage further animal and clinical MDR studies with a combination therapy consisting of NFV and chemotherapeutic drugs.

The emerging role of microRNAs and long noncoding RNAs in drug resistance of hepatocellular carcinoma

“…Adriamycin could diffuse into the nucleus of HCC cells, interact with DNA and eventually induce apoptosis. There are some known mechanisms for development of adriamycin resistance of HCC [68][69][70][71][72][73][74][75]. Multiple miRNAs have been reported to be involved in adriamycin resistance in HCC ( Table 2).…”

The Anti-Cancer Properties of the HIV Protease Inhibitor Nelfinavir

“…Liu and colleagues corroborated nelfinavir-mediated depolarization of the mitochondrial membrane in doxorubicin-resistant CML cells, which resulted in a loss of adenosine triphosphate (ATP)-suggesting induction of metabolic stress. The authors further observed an increase in ROS level during the combination of suboptimal doses of doxorubicin and nelfinavir in doxorubicin-resistant CML cells [45]. Xiang et al showed enhanced intracellular and mitochondrial ROS production in cervical cancer cells; nelfinavir-mediated cellular apoptosis was rescued by the addition of antioxidant NAC and a mitochondria-targeting superoxide and alkyl-radical scavenger Mito-TEMPO, which indicates the role of oxidative stress in nelfinavir-induced cytotoxicity [33].…”

“…However, decreased phosphorylation of ERK in cancer cells is not a universal response to nelfinavir treatment, as nelfinavir did not downregulate ERK phosphorylation in NSCLC [47], pancreatic cancer [101], and pituitary adenoma [105]. Downregulation of phospho-ERK in response to nelfinavir was further observed during combination with doxorubicin in doxorubicin-resistant chronic myeloid leukemia cells [45], and with bortezomib against MM cells [73]. Nelfinavir sensitized BRAF-mutated melanoma cells to MEK inhibitors and BRAF inhibitors via SMAD-mediated downregulation of PAX and MITF, and decreased phosphorylation of ERK during combination with inhibitors of MEK or BRAF [113].…”

“…In pediatric leukemia cells treated with nelfinavir, PARP cleavage was associated with the cleavage of upstream apoptosis initiator caspase-9 [44]. Liu et al observed a resensitization of doxorubicin-resistant chronic myeloid leukemia (CML) cells during co-treatment of suboptimal doses of nelfinavir with doxorubicin, which resulted in increased apoptosis associated to caspase-3 cleavage, increased proapoptotic protein Bax, and decreased anti-apoptotic protein Bcl-2 [45]. In castration-resistant prostate cancer cells, nelfinavir did not activate caspase-3 at low doses; however, in combination with docetaxel and curcumin, caspase-3 was activated, which resulted in DNA fragmentation and cleavage of PARP [46].…”

The effects and mechanisms of aloe‐emodin on reversing adriamycin‐induced resistance of MCF‐7/ADR cells

“…P-gp transported one molecule of the substrate across the membrane, requiring two molecules of ATP to provide energy (Robey et al, 2018). If the energy supply was insufficient, the efflux function of P-gp would be inhibited (Liu et al, 2018). Therefore, the content of intracellular ATP was measured to evaluate the effect of AE on P-gp function.…”