Profiling SLCO and SLC22 genes in the NCI-60 cancer cell lines to identify drug uptake transporters

Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA.
Molecular Cancer Therapeutics (Impact Factor: 5.68). 10/2008; 7(9):3081-91. DOI: 10.1158/1535-7163.MCT-08-0539
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Molecular and pharmacologic profiling of the NCI-60 cell panel offers the possibility of identifying pathways involved in drug resistance or sensitivity. Of these, decreased uptake of anticancer drugs mediated by efflux transporters represents one of the best studied mechanisms. Previous studies have also shown that uptake transporters can influence cytotoxicity by altering the cellular uptake of anticancer drugs. Using quantitative real-time PCR, we measured the mRNA expression of two solute carrier (SLC) families, the organic cation/zwitterion transporters (SLC22 family) and the organic anion transporters (SLCO family), totaling 23 genes in normal tissues and the NCI-60 cell panel. By correlating the mRNA expression pattern of the SLCO and SLC22 family member gene products with the growth-inhibitory profiles of 1,429 anticancer drugs and drug candidate compounds tested on the NCI-60 cell lines, we identified SLC proteins that are likely to play a dominant role in drug sensitivity. To substantiate some of the SLC-drug pairs for which the SLC member was predicted to be sensitizing, follow-up experiments were performed using engineered and characterized cell lines overexpressing SLC22A4 (OCTN1). As predicted by the statistical correlations, expression of SLC22A4 resulted in increased cellular uptake and heightened sensitivity to mitoxantrone and doxorubicin. Our results indicate that the gene expression database can be used to identify SLCO and SLC22 family members that confer sensitivity to cancer cells.

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Available from: Matthew D Hall, Feb 28, 2014
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    • "Among the SLCO family members, SLCO5A1 is the only gene which is located on chromosome 8 (8q13.3). High mRNA levels were detected in the brain, heart, skeletal muscle, and ovary [16]. SLCO5A1 was observed in human bone tumors, in prostate cancer [17] and in normal and cancerous breast tissue [18]. "
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    ABSTRACT: Organic anion transporting polypeptides (OATP/SLCO) have been identified to mediate the uptake of a broad range of mainly amphipathic molecules. Human OATP5A1 was found to be expressed in the epithelium of many cancerous and non-cancerous tissues throughout the body but protein characterization and functional analysis have not yet been performed. This study focused on the biochemical characterization of OATP5A1 using Xenopus laevis oocytes and Flp-In T-REx-HeLa cells providing evidence regarding a possible OATP5A1 function. SLCO5A1 is highly expressed in mature dendritic cells compared to immature dendritic cells (∼6.5-fold) and SLCO5A1 expression correlates with the differentiation status of primary blood cells. A core- and complex- N-glycosylated polypeptide monomer of ∼105 kDa and ∼130 kDa could be localized in intracellular membranes and on the plasma membrane, respectively. Inducible expression of SLCO5A1 in HeLa cells led to an inhibitory effect of ∼20% after 96 h on cell proliferation. Gene expression profiling with these cells identified immunologically relevant genes (e.g. CCL20) and genes implicated in developmental processes (e.g. TGM2). A single nucleotide polymorphism leading to the exchange of amino acid 33 (L→F) revealed no differences regarding protein expression and function. In conclusion, we provide evidence that OATP5A1 might be a non-classical OATP family member which is involved in biological processes that require the reorganization of the cell shape, such as differentiation and migration.
    PLoS ONE 12/2013; 8(12):e83257. DOI:10.1371/journal.pone.0083257 · 3.23 Impact Factor
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    • "However, recent results indicate that at least the two OATP isoforms primarily expressed in the liver, namely, OATP1B1 and OATP1B3, are electrogenic transporters, although their activity may be strongly affected under circumstances of abnormal variations of local pH [13]. This is pharmacologically important because tumour environment is often acidic and both transporters can be the gate for the entrance in liver tumours of many cytostatic drugs [14], such as irinotecan and its active metabolite, 7-ethyl-10-hydroxycamptothecin (SN-38) [15], flavopiridol [16], methotrexate [17], paclitaxel [18], and several bile acid-cisplatin conjugates (BAMET) [19]. OATP2B1 is expressed in territories other than liver, where this transporter is involved in the uptake of different drugs. "
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    ABSTRACT: The liver plays a crucial role in the detoxification of drugs used in the treatment of many diseases. The liver itself is the target for drugs aimed to modify its function or to treat infections and tumours affecting this organ. Both detoxification and pharmacological processes occurring in the liver require the uptake of the drug by hepatic cells and, in some cases, the elimination into bile. These steps have been classified as detoxification phase 0 and phase III, respectively. Since most drugs cannot cross the plasma membrane by simple diffusion, the involvement of transporters is mandatory. Several members of the superfamilies of solute carriers (SLC) and ATP-binding cassette (ABC) proteins, with a minor participation of other families of transporters, account for the uptake and efflux, respectively, of endobiotic and xenobiotic compounds across the basolateral and apical membranes of hepatocytes and cholangiocytes. These transporters are also involved in the sensitivity and refractoriness to the pharmacological treatment of liver tumours. An additional interesting aspect of the role of plasma membrane transporters in liver pharmacology regards the promiscuity of many of these carriers, which accounts for a variety of drug-drug, endogenous substances-drug and food components-drug interactions with clinical relevance.
    10/2012; 2012(8):428139. DOI:10.6064/2012/428139
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    • "Dasatinib N/A ABCB1, ABCC4, ABCG2 (Dohse et al., 2010) Daunorubicin SLC22A4 (OCTN1) ABCB1 (Kartner et al., 1983; Okabe et al., 2008 "
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    ABSTRACT: Members of the solute carrier family of transporters are responsible for the cellular uptake of a broad range of endogenous compounds and xenobiotics in multiple tissues. Several of these solute carriers are known to be expressed in cancer cells or cancer cell lines, and decreased cellular uptake of drugs potentially contributes to the development of resistance. As result, the expression levels of these proteins in humans have important consequences for an individual's susceptibility to certain drug-induced side effects, interactions, and treatment efficacy. In this review article, we provide an update of this rapidly emerging field, with specific emphasis on the direct contribution of solute carriers to anticancer drug uptake in tumors, the role of these carriers in regulation of anticancer drug disposition, and recent advances in attempts to evaluate these proteins as therapeutic targets.
    Drug resistance updates: reviews and commentaries in antimicrobial and anticancer chemotherapy 03/2012; 15(1-2):5-20. DOI:10.1016/j.drup.2012.01.009 · 9.12 Impact Factor
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