The Reversal Effects of 3-Bromopyruvate on Multidrug Resistance In Vitro and In Vivo Derived from Human Breast MCF-7/ADR Cells

Abstract

PurposeP-glycoprotein mediated efflux is one of the main mechanisms for multidrug resistance in cancers, and 3-Bromopyruvate acts as a promising multidrug resistance reversal compound in our study. To test the ability of 3-Bromopyruvate to overcome P-glycoprotein-mediated multidrug resistance and to explore its mechanisms of multidrug resistance reversal in MCF-7/ADR cells, we evaluate the in vitro and in vivo modulatory activity of this compound.MethodsThe in vitro and in vivo activity was determined using the MTT assay and human breast cancer xenograft models. The gene and protein expression of P-glycoprotein were determined using real-time polymerase chain reaction and the Western blotting technique, respectively. ABCB-1 bioactivity was tested by fluorescence microscopy, multi-mode microplate reader, and flow cytometry. The intracellular levels of ATP, HK-II, and ATPase activity were based on an assay kit according to the manufacturer’s instructions.Results3-Bromopyruvate treatment led to marked decreases in the IC50 values of selected chemotherapeutic drugs [e.g., doxorubicin (283 folds), paclitaxel (85 folds), daunorubicin (201 folds), and epirubicin (171 folds)] in MCF-7/ADR cells. 3-Bromopyruvate was found also to potentiate significantly the antitumor activity of epirubicin against MCF-7/ADR xenografts. The intracellular level of ATP decreased 44%, 46% in the presence of 12.5.25 µM 3-Bromopyruvate, whereas the accumulation of rhodamine 123 and epirubicin (two typical P-glycoprotein substrates) in cells was significantly increased. Furthermore, we found that the mRNA and the total protein level of P-glycoprotein were slightly altered by 3-Bromopyruvate. Moreover, the ATPase activity was significantly inhibited when 3-Bromopyruvate was applied.ConclusionWe demonstrated that 3-Bromopyruvate can reverse P-glycoprotein-mediated efflux in MCF-7/ADR cells. Multidrug resistance reversal by 3-Bromopyruvate occurred through at least three approaches, namely, a decrease in the intracellular level of ATP and HK-II bioactivity, the inhibition of ATPase activity, and the slight decrease in P-glycoprotein expression in MCF-7/ADR cells.

Metabolic Suppression of a Drug‐Resistant Subpopulation in Cancer Spheroid Cells

“…The efficiency of glycolytic inhibitors in overcoming drug efflux was reported in multiple 2-D-grown cancer cell types [Kaplan et al, 1990;Maschek et al, 2004], including the MCF-7 breast cancer cell line used in the present work [Wu et al, 2014]. Therefore, we compared effects of glycolytic inhibition on MDR in 2-D and 3-D cell culture systems.…”

3‐Bromopyruvate as a potent anticancer therapy in honor and memory of the late Professor André Goffeau

“…As one of 3 BP 's characteristics, it exhibits synergistic effects with cytostatic agents that are substrates for ABC superfamily transporters. The ATP deficit due to the action of 3 BP prevents full operation of the efflux pumps, which are not able to pump out of the cells large quantities of the following agents: cisplatin, doxorubicin, 5‐fluorouracil, paclitaxel, epirubicin, daunorubicin, and mitoxantrone (Rello et al, ; Sánchez‐Aragó & Cuezva, ; Wu et al, ). Moreover, previous studies showed that 3 BP acts synergistically with compounds, which are GSH‐depleting agents such as buthionine sulfoximine, methionine sulfoximine, and acetaminophen (El Sayed et al, ; El Sayed, Baghdadi, Almaramhy, Ayat, & Donki, ; Garbutcheon‐Singh, Harper, & Aldrich‐Wright, ; Lee et al, ; Lis, Dyląg, et al, ; Majkowska‐Skrobek et al, ; Niedźwiecka et al, ).…”

“…In addition to 3 BP 's effectiveness as an anticancer agent, its various utilizations continues to expand for other conditions, that is, antifibrotic effects in liver (Karthikeyan et al, ), immune modulation (Jung, Lee, Park, Quach, & Lee, ; Yadav et al, ), antiangiogenesis (El‐Sayed et al, ), reversal of multidrug resistance (Wu et al, ), infection (parasite, fungi, bacteria, and virus; Barnard et al, ; Niedźwiecka et al, ), rheumatoid arthritis (Garcia‐Carbonell et al, ), seizure (Lee et al, ), keloid scar formation (Vincent, ), and pulmonary arterial hypertension (Chen, Wang, Lai, Cai, & Yuan, ; Zhang et al, ).…”

“…The results obtained may provide the basis for further research focused on designing new drugs and strategies for the treatment of cancer, cryptococcosis, and protothecosis. It is also known that 3 BP demonstrates synergistic activity in combination with other agents, for example, cisplatin, doxorubicin, 5‐fluorouracil, paclitaxel, epirubicin, daunorubicin, and mitoxantrone (Nakano et al, ; Sánchez‐Aragó & Cuezva, ; Wu et al, ).…”

Hepatotoxicity and nephrotoxicity of 3-bromopyruvate in mice

“…As a glycolytic inhibitor, "3BP is a fast acting, promising, powerful, specific and effective anti-cancer agent", said by Pedersen 3 . Also 3BP has been identified to inhibit tumor cells proliferation as a proposed widely used anti-cancer agent 4 , such as in breast cancer cells 5,6 , liver cancer cells 7,8 , lymphoma cells 9 , lung cancer cells 10 , pancreatic cancer cells 11,12 , gastric cancer 2,13 , colorectal cancer cells 14 . Those significant anti-tumor effects of 3BP has made it getting more and more attention 15,16 .…”