Functional investigation of a highly conserved aspartate residue in the anti -cancer drug efflux transport protein MRP1

2018 ◽  
Author(s):  
Graeme Mullins
Keyword(s):  
Abc Transporters ◽  
Double Mutant ◽  
Transmembrane Helix ◽  
Salt Bridge ◽  
Multidrug Resistant ◽  
Site Directed Mutagenesis ◽  
Cancer Drug ◽  
Wide Range ◽  
Anti Cancer ◽  
Membrane Spanning

Multidrug Resistant Protein 1 (MRP1 or ABCC1) belongs to a subclass of ATP-binding cassette (ABC) transporters that export a wide range of metabolites and xenobiotics across the plasma membrane. Increased expression of MRP1 in cancer cells enhances efflux of many anti-cancer agents, giving rise to multidrug resistant tumours. The purpose of this study was to investigate the function of an aspartate (Asp) amino acid that is highly conserved in all MRP-related proteins by mutating it and determining the consequences of doing so. Asp430 lies at the interface of the cytoplasm and a transmembrane helix in the first membrane-spanning domain of MRP1. Previous studies have shown that when Asp430 is mutated, the protein becomes unstable and is degraded.Because this Asp430 is highly conserved in many MRP-related ABC transporters and because structural homology models of human MRP1 predict that Asp430 is in close proximity to Arg433, we hypothesized that a salt bridge between these two a mino acids could be essential for proper folding and stability of the protein during its biosynthesis. Using site -directed mutagenesis, these two amino acids were interchanged to probe the existence of such an interaction. Thus a double mutant where Asp430 was mutated to Arg, and Arg433 was mutated to Asp was created, and the resultant mutant protein (D430R/R433D) was tested for its ability to be detected in mammalian cells by gel electrophoresis and immunoblotting. Our results show differences between the migration patterns of double and single mutants that are compatible with differences in the glycosylation levels of MRP1. However the fact that D430R and the R433D mutants don’t share the same migration pattern, together with the variation in migration bet ween D430 wild type and Supported by CIHR MOP-10519the double mutant D430R/R433D indicate that the possibility of a salt bridge can be discarded.Supported by CIHR MOP-10519

Abstract 5045: Overexpression of ABC transporters through HGF/c-MET activation results in anti-cancer drug resistance in SCLC

2016 ◽  
Author(s):  
Yoshitaka Yagi ◽  
Hiroaki Ozasa ◽  
Takahiro Tsuji ◽  
Yuichi Sakamori ◽  
Takeshi Nomizo ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Abc Transporters ◽  
Cancer Drug ◽  
Cancer Drug Resistance ◽  
Anti Cancer

scholarly journals The study of drug resistance properties of ABCG2 (ATP-binding cassette G2) in contact with thymoquinone, gallic acid, and hesperetin antioxidants

2019 ◽  
Vol 8 (2) ◽  
pp. 108-113 ◽  
Author(s):  
Javad Saffari_Chaleshtori ◽  
Sayed Mohammad Shafiee ◽  
Keihan Ghatreh-Samani ◽  
Narges Jalilian
Keyword(s):  
Binding Site ◽  
Gallic Acid ◽  
Abc Transporters ◽  
Data Bank ◽  
Effective Concentration ◽  
Cancer Drug ◽  
Atp Binding ◽  
Anti Cancer ◽  
Abcg2 Protein ◽  
Atp Binding Cassette

Introduction: ATP-binding cassette (ABC) transporters are a group of intra membrane proteins that play key roles in the transmission and exchange of vital compounds on both sides of the membrane. These proteins can specially transport anti-cancer drugs out of cancer cells. ABCG2 is a member of this family that is extremely expressed in many cancers. This study, aims to evaluate the binding affinity of three antioxidants thymoquinone (TQ), gallic acid (GA), and hesperetin (HP) to ABCG2 compared with an anti-cancer drug, mitoxantrone (Mit), to export cells. Methods: The PDB file of ABCG2 was obtained from the protein data bank server (http://www.rcsb.org) with ID: 5NJ3. After 200 stages of molecular docking running on ABCG2 protein in AutoDock v.4.2 software, the amino acids involved in the binding site of each compound were identified using the LigPlot+ software. Results: HP had the lowest (-6.36 kcal/mol) and GA had the highest (-3.93 kcal/mol) binding energy in comparison with Mit (-0.06 kcal/mol) for binding to ABCG2. Effective concentration required to perform the reaction between ABCG2 was higher in GA (1.31 mM) than TQ (42.69 μM) and HP (21.74 μM). GA, HP, and TQ formed 17, 18, and 22 hydrogen and hydrophobic bonds at the binding site of ABCG2. Conclusion: It seems that GA has the lowest affinity to make contact with ABCG2 binding site. So, GA tends to remain in the cell but TQ and HP tend to leave the cell easily via ABCG2 transporter.

Effect of Metformin Nanoparticle-Mediated Thioredoxin Interacting Protein Expression on Oxaliplatin-Induced Peripheral Neuralgia

2020 ◽  
Vol 20 (10) ◽  
pp. 6123-6132 ◽  
Author(s):  
Yun Liu ◽  
Guangquan Zhou ◽  
Nenggui Xu
Keyword(s):  
Neuropathic Pain ◽  
Sciatic Nerve ◽  
Peripheral Nerves ◽  
Pain Perception ◽  
Thermal Sensitivity ◽  
Cancer Drug ◽  
Pain Model ◽  
Chemotherapy Drugs ◽  
Wide Range ◽  
Anti Cancer

Oxaliplatin (LOHP) is an approved anti-cancer drug that often accumulates in the peripheral nerves during the treatment of cancer. Metformin is a prescription drug with a wide range of pharmacological effects, which can augment the anti-cancer efficacy of chemotherapy drugs. Recent studies suggest that metformin is a potential drug that can relieve oxaliplatin induced neuralgia, and autophagy plays an important role in it. This study aims at exploring the effect and mechanism of action of metformin on oxaliplatin-induced peripheral neuropathic pain. To explore the underlying mechanism of metformin on peripheral nerves, neuropathic pain model was developed by intraperitoneal injection of oxaliplatin into mice. Metformin nanoparticles encapsulated by PLGA were used to intervene the pain in the model mice. RT-qPCR and immunoblotting were used to detect the effects of metformin on the expression of TXNIP and autophagy associated genes-BECN1 and LC3B in the sciatic nerves of pain model mice. Hematoxylin and eosin staining were used to detect the pathological changes in the sciatic nerve. Flow cytometry and Annexin V-FITC/PI apoptosis detection kit were used to detect the apoptosis of sciatic nerve cells. The effect of metformin on the pain perception of mice was detected by thermal and mechanical stimulation experiments. The results showed that the expression of TXNIP and autophagy related indexes-BECN1 and LC3B in sciatic nerve decreased significantly after oxaliplatin treatment. However, metformin intervention resulted in significant up-regulation of TXNIP and autophagy related indexes, and augmented the threshold of thermal sensitivity and mechanical tingling. Thus, our study has identified TXNIP as a novel target for oxaliplatin induced peripheral nerve pain. We have shown that oxaliplatin inhibits TXNIP expression, regulates autophagy, thereby affecting neuralgia. In contrast, metformin promotes the expression of TXNIP and autophagy of cells thereby inhibiting neural sensitivity and thus results in pain relief.

Rifampicin enhances anti-cancer drug accumulation and activity in multidrug-resistant cells

1995 ◽  
Vol 49 (9) ◽  
pp. 1255-1260 ◽  
Author(s):  
Olivier Fardel ◽  
Valérie Lecureur ◽  
Pascal Loyer ◽  
André Guillouzo
Keyword(s):  
Multidrug Resistant ◽  
Drug Accumulation ◽  
Cancer Drug ◽  
Anti Cancer ◽  
Resistant Cells

Ursolic Acid in Cancer Treatment and Metastatic Chemoprevention: From Synthesized Derivatives to Nanoformulations in Preclinical Studies

2019 ◽  
Vol 19 (4) ◽  
pp. 245-256 ◽  
Author(s):  
Junjie Zou ◽  
Juanfang Lin ◽  
Chao Li ◽  
Ruirui Zhao ◽  
Lulu Fan ◽  
...  
Keyword(s):  
Ursolic Acid ◽  
Cancer Metastasis ◽  
Biological Activities ◽  
Inhibitory Effect ◽  
Cancer Drug ◽  
Therapeutic Effects ◽  
Candidate Drug ◽  
Wide Range ◽  
Anti Cancer ◽  
Tumor Growth And Metastasis

Background:Cancer metastasis has emerged as a major public health threat that causes majority of cancer fatalities. Traditional chemotherapeutics have been effective in the past but suffer from low therapeutic efficiency and harmful side-effects. Recently, it has been reported ursolic acid (UA), one of the naturally abundant pentacyclic triterpenes, possesses a wide range of biological activities including anti-inflammatory, anti-atherosclerotic, and anti-cancer properties. More importantly, UA has the features of low toxicity, liver protection and the potential of anti-cancer metastasis.Objective:This article aimed at reviewing the great potential of UA used as a candidate drug in the field of cancer therapy relating to suppression of tumor initiation, progression and metastasis.Methods:Selective searches were conducted in Pubmed, Google Scholar and Web of Science using the keywords and subheadings from database inception to December 2017. Systemic reviews are summarized here.Results:UA has exhibited chemopreventive and therapeutic effects of cancer mainly through inducing apoptosis, inhibiting cell proliferation, preventing tumor angiogenesis and metastatic. UA nanoformulations could enhance the solubility and bioavailability of UA as well as exhibit better inhibitory effect on tumor growth and metastasis.Conclusion:The information presented in this article can provide useful references for further studies on making UA a promising anti-cancer drug, especially as a prophylactic metastatic agent for clinical applications.

scholarly journals Cellular Analysis and Chemotherapeutic Potential of a Bi-Functionalized Halloysite Nanotube

Pharmaceutics ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 962
Author(s):  
Yangyang Luo ◽  
Ahmed Humayun ◽  
Teresa A. Murray ◽  
Benjamin S. Kemp ◽  
Antwine McFarland ◽  
...  
Keyword(s):  
Cancer Cells ◽  
Drug Carrier ◽  
Drug Loading ◽  
Cancer Drug ◽  
Intracellular Location ◽  
Cellular Analysis ◽  
Osteoblast Cell Line ◽  
Wide Range ◽  
Anti Cancer ◽  
Target Cancer

The surface of halloysite nanotubes (HNTs) was bifunctionalized with two ligands—folic acid and a fluorochrome. In tandem, this combination should selectively target cancer cells and provide a means for imaging the nanoparticle. Modified bi-functionalized HNTs (bi-HNTs) were then doped with the anti-cancer drug methotrexate. bi-HNTs were characterized and subjected to in vitro tests to assess cellular growth and changes in cellular behavior in three cell lines—colon cancer, osteosarcoma, and a pre-osteoblast cell line (MC3T3-E1). Cell viability, proliferation, and cell uptake efficiency were assessed. The bi-HNTs showed cytocompatibility at a wide range of concentrations. Compared with regular-sized HNTs, reduced HNTs (~6 microns) were taken up by cells in more significant amounts, but increased cytotoxicity lead to apoptosis. Multi-photon images confirmed the intracellular location of bi-HNTs, and the method of cell entry was mainly through caveolae-mediated endocytosis. The bi-HNTs showed a high drug loading efficiency with methotrexate and a prolonged period of release. Most importantly, bi-HNTs were designed as a drug carrier to target cancer cells specifically, and imaging data shows that non-cancerous cells were unaffected after exposure to MTX-doped bi-HNTs. All data provide support for our nanoparticle design as a mechanism to selectively target cancer cells and significantly reduce the side-effects caused by off-targeting of anti-cancer drugs.

scholarly journals Rewiring of lactate-IL-1β auto-regulatory loop with Clock-Bmal1: A feed-forward circuit in glioma

2021 ◽  
Author(s):  
Pruthvi Gowda ◽  
Kirti Lathoria ◽  
Shalini Sharma ◽  
Shruti Patrick ◽  
Sonia B. Umdor ◽  
...  
Keyword(s):  
Site Directed Mutagenesis ◽  
Luciferase Reporter ◽  
Cancer Drug ◽  
Cancer Etiology ◽  
Autocrine Loop ◽  
Expression Of Genes ◽  
Anti Cancer ◽  
E Box ◽  
Cervical Cancer Cells ◽  
Regulatory Loop

De-synchronized circadian rhythm in tumors is coincident with aberrant inflammation and dysregulated metabolism. As their inter-relationship in cancer etiology is largely unknown, we investigated the link between the three in glioma. Tumor metabolite lactate- mediated increase in pro-inflammatory cytokine IL-1β was concomitant with elevated levels of core circadian regulators Clock and Bmal1. siRNA mediated knockdown of Bmal1 and Clock decreased (i) LDHA and IL-1β levels and (ii) release of lactate and pro-inflammatory cytokines. Lactate mediated deacetylation of Bmal1 and its interaction with Clock, regulate IL-1β levels and vice versa. Site-directed mutagenesis and luciferase reporter assay indicated the functionality of E-box sites on LDHA and IL-1β promoters. ChIP-re-ChIP revealed that lactate-IL-1β crosstalk positively affects co-recruitment of Clock-Bmal1 to these E-box sites. Clock-Bmal1 enrichment was accompanied by decreased H3K9me3, and increased H3K9ac and RNA pol II occupancy. Lactate-IL-1β-Clock (LIC) loop positively regulated expression of genes associated with cell cycle, DNA damage and cytoskeletal organization involved in glioma progression. TCGA data analysis suggested the presence of lactate- IL-1β-crosstalk in other cancers. The responsiveness of stomach and cervical cancer cells to lactate inhibition followed the same trend exhibited by glioma cells. In addition, components of LIC loop were found to be correlated with (i) patient survival, (ii) clinically actionable genes, and (iii) anti-cancer drug sensitivity. Our findings provide evidence for a potential cancer-specific axis wiring of IL-1β and LDHA through Clock -Bmal1, the outcome of which is to fuel an IL-1β-lactate autocrine loop that drives pro-inflammatory and oncogenic signals.

scholarly journals Tannic Acid-Lung Fluid Assemblies Promote Interaction and Delivery of Drugs to Lung Cancer Cells

Pharmaceutics ◽  
2018 ◽  
Vol 10 (3) ◽  
pp. 111 ◽  
Author(s):  
Elham Hatami ◽  
Prashanth Nagesh ◽  
Pallabita Chowdhury ◽  
Subhash Chauhan ◽  
Meena Jaggi ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Tannic Acid ◽  
Biological Activities ◽  
The United States ◽  
Water Soluble ◽  
Cancer Drug ◽  
Major Barrier ◽  
Lung Fluid ◽  
Wide Range ◽  
Anti Cancer

Lung cancer (LC) is one of the leading causes of death in both men and women in the United States. Tannic acid (TA), a water-soluble polyphenol, exhibits a wide range of biological activities. TA has received much attention as a promising compound in the biomaterial and drug delivery fields. Lung fluid (LF) is a major barrier for distribution of drugs to the lungs. Therefore, the purpose of this study was to examine TA interaction with LF for effective delivery of anti-cancer drug molecules via pulmonary delivery. The extent of adsorption of LF proteins by TA was revealed by fluorescence quenching in fluorescence spectroscopy. The presence of LF in TA-LF complexes was noticed by the presence of protein peaks at 1653 cm−1. Both protein dot and SDS-PAGE analysis confirmed LF protein complexation at all TA concentrations employed. A stable particle TA-LF complex formation was observed through transmission electron microscopy (TEM) analysis. The complexation pattern measured by dynamic light scattering (DLS) indicated that it varies depending on the pH of the solutions. The degree of LF presence in TA-LF complexes signifies its interactive behavior in LC cell lines. Such superior interaction offered an enhanced anti-cancer activity of drugs encapsulated in TA-LF complex nanoformulations. Our results indicate that TA binds to LF and forms self-assemblies, which profoundly enhance interaction with LC cells. This study demonstrated that TA is a novel carrier for pharmaceutical drugs such as gemcitabine, carboplatin, and irinotecan.

Deactivation of anti-cancer drug letrozole to a carbinol metabolite by polymorphic cytochrome P450 2A6 in human liver microsomes

Xenobiotica ◽  
2009 ◽  
Vol 00 (00) ◽  
pp. 090901052053001-8
Author(s):  
K. Murai ◽  
H. Yamazaki ◽  
K. Nakagawa ◽  
R. Kawai ◽  
T. Kamataki
Keyword(s):  
Cytochrome P450 ◽  
Human Liver ◽  
Liver Microsomes ◽  
Human Liver Microsomes ◽  
Cancer Drug ◽  
Anti Cancer ◽  
Cytochrome P450 2A6
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