Lactate-induced MRP1 expression contributes to metabolism-based etoposide resistance in non-small cell lung cancer cells

Abstract

Background Metabolic reprogramming contributes significantly to tumor development and is tightly linked to drug resistance. The chemotherapeutic agent etoposide (VP-16) has been used clinically in the treatment of lung cancer but possess different sensitivity and efficacy towards SCLC and NSCLC. Here, we assessed the impact of etoposide on glycolytic metabolism in SCLC and NSCLC cell lines and investigated the role of metabolic rewiring in mediating etoposide resistance. Methods glycolytic differences of drug-treated cancer cells were determined by extracellular acidification rate (ECAR), glucose consumption, lactate production and western blot. DNA damage was evaluated by the comet assay and western blot. Chemoresistant cancer cells were analyzed by viability, apoptosis and western blot. Chromatin immunoprecipitation (ChIP) was used for analysis of DNA-protein interaction. Results Here we showed that exposure to chemotherapeutic drug etoposide induces an exacerbation of ROS production which activates HIF-1α-mediated the metabolic reprogramming toward increased glycolysis and lactate production in non-small cell lung cancer (NSCLC). We identified lactic acidosis as the key that confers multidrug resistance through upregulation of multidrug resistance-associated protein 1 (MRP1, encoded by ABCC1), a member of ATP-binding cassette (ABC) transporter family. Mechanistically, lactic acid coordinates TGF-β1/Snail and TAZ/AP-1 pathway to induce formation of Snail/TAZ/AP-1 complex at the MRP1/ABCC1 promoter. Induction of MRP1 expression inhibits genotoxic and apoptotic effects of chemotherapeutic drugs by increasing drug efflux. Furthermore, titration of lactic acid with NaHCO3 was sufficient to overcome resistance. Conclusions The chemotherapeutic drug etoposide induces the shift toward aerobic glycolysis in the NSCLC rather than SCLC cell lines. The increased lactic acid in extracellular environment plays important role in etoposide resistance through upregulation of MRP expression. These data provide first evidence for the increased lactate production, upon drug treatment, contributes to adaptive resistance in NSCLC and reveal potential vulnerabilities of lactate metabolism and/or pathway suitable for therapeutic targeting. Graphical abstract

The Mechanism of Warburg Effect-Induced Chemoresistance in Cancer

“…This phenomenon is called the "ion trapping mechanism" (227)(228)(229)(230)(231). Meanwhile, extracellular acidosis facilitates P-gp, ABC subfamily B member 1 (ABCB1) and 2 (ABCB2) transporter, and intracellular acidic vesicles to remove drugs out of cancer cells, further inducing chemoresistance (224,(232)(233)(234)(235). (2) Extracellular acidosis promotes chemoresistance signaling pathways by activating related proteins.…”

“…(2) Extracellular acidosis promotes chemoresistance signaling pathways by activating related proteins. For example, mild acidic stress not only facilitates unfolded protein response (UPR) but also triggers an adaptive UPR with progressive increase in glucose regulatory protein 78 expression, which reduces the cleavage of caspase 7 to induce sunitinib resistance in oral squamous cancer (212,224); extracellular lactate functions as an agonist for G protein-coupled receptor 81 (GPR81) and promotes GPR81 upregulation of the PI3K/AKT/mTOR pathway to inhibit apoptosis, promote stem cell phenotype, inhibit immune response, and induce etoposide resistance in non-small-cell lung cancer (234,236). (3) Lactate inhibits immune response in different ways: ① lactate directly inhibits the cytotoxicity of perforin and granzyme; ② high extracellular lacttate levels lead to the accumulation of endogenous lacttate in T cells, thereby reducing the secretion of pro-inflammatory cytokine; ③ lacttate indirectly weakens natural killer (NK) cell function by recruiting monocyte-derived dendritic cells (225); and (4) extracellular acidosis not only has synergistic effect with hypoxia but also facilitates glycolysis of stromal cells to produce lactate for fueling cancer cells, ensuring survival and proliferation and preventing apoptosis (166).…”

The role of non‐coding RNAs in drug resistance of oral squamous cell carcinoma and therapeutic potential

“…MRP1 , on the other hand, outputs both endobiotics and xenobiotics, thereby affecting physiological processes beyond drug distribution [ 83 , 84 ]. The expression level of MRP1 is known to be significantly upregulated in several drug‐resistant diseases, including non‐small cell lung cancer [ 85 ] and epilepsy [ 86 ]. In CDDP‐resistant A549 cells, expression levels of lncRNA MALAT1 and MRP1 were upregulated compared to A549 cells.…”

Stimulation of ROS Generation by Extract of Warburgia ugandensis Leading to G0/G1 Cell Cycle Arrest and Antiproliferation in A549 Cells

“…Natural products derived from medicinal plants have been used for anticancer therapy based on ROS-induced cell death [54][55][56]. Stimulating ROS generation is also a widely accepted mechanism that is manipulated by a variety of anti-NSCLC chemotherapeutical drugs, including cisplatin, etoposide, doxorubicin, docetaxel, and paclitaxel [57][58][59][60][61][62]. Compared to normal cells, cancer cells usually contain a higher basal level of ROS, which makes cancer cells are more vulnerable to damage by further ROS insults.…”