Acidosis induces multi-drug resistance in rat prostate cancer cells (AT1) in vitro and in vivo by increasing the activity of the p-glycoprotein via activation of p38
Christoph Sauvant
Physiologisches Institut, Universität Würzburg, Würzburg, Germany
C. Sauvant and M. Nowak contributed equally to this work and are both to be considered as first authors.
Search for more papers by this authorMartin Nowak
Institut für Physiologie und Pathophysiologie, Universität Mainz, Mainz, Germany
C. Sauvant and M. Nowak contributed equally to this work and are both to be considered as first authors.
Search for more papers by this authorClaudia Wirth
Physiologisches Institut, Universität Würzburg, Würzburg, Germany
Search for more papers by this authorBettina Schneider
Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Halle, Germany
Search for more papers by this authorAnne Riemann
Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Halle, Germany
Search for more papers by this authorCorresponding Author
Michael Gekle
Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Halle, Germany
M. Gekle and O. Thews contributed equally to this work and are both to be considered as senior authors.
Fax: +49-345-557-4019.
Michael Gekle, Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Magdeburger Strasse 6, 06112 Halle (Saale), Germany
Oliver Thews, Institut für Physiologie und Pathophysiologie, Universität Mainz, Duesbergweg 6, 55099 Mainz, Germany
Search for more papers by this authorCorresponding Author
Oliver Thews
Institut für Physiologie und Pathophysiologie, Universität Mainz, Mainz, Germany
M. Gekle and O. Thews contributed equally to this work and are both to be considered as senior authors.
Fax: +496-131-392-4543.
Michael Gekle, Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Magdeburger Strasse 6, 06112 Halle (Saale), Germany
Oliver Thews, Institut für Physiologie und Pathophysiologie, Universität Mainz, Duesbergweg 6, 55099 Mainz, Germany
Search for more papers by this authorChristoph Sauvant
Physiologisches Institut, Universität Würzburg, Würzburg, Germany
C. Sauvant and M. Nowak contributed equally to this work and are both to be considered as first authors.
Search for more papers by this authorMartin Nowak
Institut für Physiologie und Pathophysiologie, Universität Mainz, Mainz, Germany
C. Sauvant and M. Nowak contributed equally to this work and are both to be considered as first authors.
Search for more papers by this authorClaudia Wirth
Physiologisches Institut, Universität Würzburg, Würzburg, Germany
Search for more papers by this authorBettina Schneider
Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Halle, Germany
Search for more papers by this authorAnne Riemann
Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Halle, Germany
Search for more papers by this authorCorresponding Author
Michael Gekle
Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Halle, Germany
M. Gekle and O. Thews contributed equally to this work and are both to be considered as senior authors.
Fax: +49-345-557-4019.
Michael Gekle, Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Magdeburger Strasse 6, 06112 Halle (Saale), Germany
Oliver Thews, Institut für Physiologie und Pathophysiologie, Universität Mainz, Duesbergweg 6, 55099 Mainz, Germany
Search for more papers by this authorCorresponding Author
Oliver Thews
Institut für Physiologie und Pathophysiologie, Universität Mainz, Mainz, Germany
M. Gekle and O. Thews contributed equally to this work and are both to be considered as senior authors.
Fax: +496-131-392-4543.
Michael Gekle, Julius-Bernstein-Institut für Physiologie, Universität Halle-Wittenberg, Magdeburger Strasse 6, 06112 Halle (Saale), Germany
Oliver Thews, Institut für Physiologie und Pathophysiologie, Universität Mainz, Duesbergweg 6, 55099 Mainz, Germany
Search for more papers by this authorM. Gekle and O. Thews contributed equally to this work and are both to be considered as senior authors.
Fax: +49-345-557-4019.
Fax: +496-131-392-4543.
Abstract
Because solid growing tumors often show hypoxia and pronounced extracellular acidosis, the aim of this study was to analyze the impact of an acidotic environment on the activity of the p-glycoprotein (pGP) and on the cellular content and cytotoxicity of the chemotherapeutic drug daunorubicin in the AT1 R-3327 Dunning prostate carcinoma cell line cultured in vitro and in vivo. In vitro, extracellular acidosis (pH 6.6) activated p38 and ERK1/2 and thereby induced daunorubicin resistance via a pronounced activation of pGP. De-novo protein synthesis was not necessary and analysis of transport kinetics indicated a fast and persistent pGP activation at pH 6.6 (when compared with 7.4). Intracellular acidification also induced daunorubicin resistance via activation of pGP, which was mediated by activation of p38 alone. In vivo, tumors were implanted subcutaneously, and the tumor pH was artificially lowered by forcing anaerobic metabolism. In vivo, the reduced extracellular pH of 6.6 was also able to induce daunorubicin resistance, which was abolished by inhibition of p38. These results suggest that pGP activity is dependent on extracellular pH in vitro and in vivo. Moreover, there is strong indication that this effect is mediated via activation of p38 in vivo. Activation of ERK is also suitable to induce pGP activity. Therefore, inhibition of p38 (and ERK) may be used to prevent acidosis induced increase in pGP activity and thereby attenuate multidrug resistance. In addition, supportive treatments reducing tumor acidosis may improve the cytotoxic effect of chemotherapeutic drugs. © 2008 Wiley-Liss, Inc.
References
- 1 Chang G. Multidrug resistance ABC transporters. FEBS Lett 2003; 555: 102–5.
- 2 Higgins CF. ABC transporters: physiology, structure and mechanism—an overview. Res Microbiol 2001; 152: 205–10.
- 3 Ambudkar SV,Kimchi-Sarfaty C,Sauna ZE,Gottesman MM. P-glycoprotein: from genomics to mechanism. Oncogene 2003; 22: 7468–85.
- 4 Higgins CF. Multiple molecular mechanisms for multidrug resistance transporters. Nature 2007; 446: 749–57.
- 5 Schinkel AH,Jonker JW. Mammalian drug efflux transporters of the ATP binding cassette (ABC) family: an overview. Adv Drug Deliv Rev 2003; 55: 3–29.
- 6 Fojo T,Bates S. Strategies for reversing drug resistance. Oncogene 2003; 22: 7512–23.
- 7 Xu RH,Pelicano H,Zhou Y,Carew JS,Feng L,Bhalla KN,Keating MJ,Huang P. Inhibition of glycolysis in cancer cells: a novel strategy to overcome drug resistance associated with mitochondrial respiratory defect and hypoxia. Cancer Res 2005; 65: 613–21.
- 8 Puddu P,Fais S,Luciani F,Gherardi G,Dupuis ML,Romagnoli G,Ramoni C,Cianfriglia M,Gessani S. Interferon-gamma up-regulates expression and activity of P-glycoprotein in human peripheral blood monocyte-derived macrophages. Lab Invest 1999; 79: 1299–309.
- 9 Yang JM,Sullivan GF,Hait WN. Regulation of the function of P-glycoprotein by epidermal growth factor through phospholipase C. Biochem Pharmacol 1997; 53: 1597–604.
- 10 Lotz C,Kelleher DK,Gassner B,Gekle M,Vaupel P,Thews O. Role of the tumor microenvironment in the activity and expression of the p-glycoprotein in human colon carcinoma cells. Oncol Rep 2007; 17: 239–44.
- 11 Thews O,Gassner B,Kelleher DK,Schwerdt G,Gekle M. Impact of extracellular acidity on the activity of p-glycoprotein and the cytotoxicity of chemotherapeutic drugs. Neoplasia 2006; 8: 143–52.
- 12 Konerding MA,Malkusch W,Klapthor B,van Ackern C,Fait E,Hill SA,Parkins C,Chaplin DJ,Presta M,Denekamp J. Evidence for characteristic vascular patterns in solid tumours: quantitative studies using corrosion casts. Br J Cancer 1999; 80: 724–32.
- 13 Vaupel P,Kallinowski F,Okunieff P. Blood flow, oxygen and nutrient supply, and metabolic microenvironment of human tumors: a review. Cancer Res 1989; 49: 6449–65.
- 14 Höckel M,Vaupel P. Tumor hypoxia: definitions and current clinical, biological, and molecular aspects. J Natl Cancer Inst 2001; 93: 266–76.
- 15 Comerford KM,Wallace TJ,Karhausen J,Louis NA,Montalto MC,Colgan SP. Hypoxia-inducible factor-1-dependent regulation of the multidrug resistance (MDR1) gene. Cancer Res 2002; 62: 3387–94.
- 16 Ledoux S,Yang R,Friedlander G,Laouari D. Glucose depletion enhances P-glycoprotein expression in hepatoma cells: role of endoplasmic reticulum stress response. Cancer Res 2003; 63: 7284–90.
- 17 Wartenberg M,Gronczynska S,Bekhite MM,Saric T,Niedermeier W,Hescheler J,Sauer H. Regulation of the multidrug resistance transporter P-glycoprotein in multicellular prostate tumor spheroids by hyperthermia and reactive oxygen species. Int J Cancer 2005; 113: 229–40.
- 18 Lee G,Piquette-Miller M. Cytokines alter the expression and activity of the multidrug resistance transporters in human hepatoma cell lines; analysis using RT-PCR and cDNA microarrays. J Pharm Sci 2003; 92: 2152–63.
- 19 Versteeg HH,Nijhuis E,van den Brink GR,Evertzen M,Pynaert GN,van Deventer SJ,Coffer PJ,Peppelenbosch MP. A new phosphospecific cell-based ELISA for p42/p44 mitogen-activated protein kinase (MAPK), p38 MAPK, protein kinase B and cAMP-response-element-binding protein. Biochem J 2000; 350: 717–22.
- 20 Sauvant C,Holzinger H,Gekle M. Proximal tubular toxicity of ochratoxin A is amplified by simultaneous inhibition of the extracellular signal-regulated kinases 1/2. J Pharmacol Exp Ther 2005; 313: 234–41.
- 21 Workman P,Twentyman P,Balkwill F,Balmain A,Chaplin DJ,Double JA,Embleton J,Newell D,Raymond R,Stables J,Stephens T,Wallace J. United Kingdom Co-ordinating Committee on Cancer Research (UKCCCR) Guidelines for the Welfare of Animals in Experimental Neoplasia (2nd edit.). Br J Cancer 1998; 77: 1–10.
- 22 Kalliomäki T,Hill RP. Effects of tumour acidification with glucose+MIBG on the spontaneous metastatic potential of two murine cell lines. Br J Cancer 2004; 90: 1842–9.
- 23 Vaupel P,Schlenger K,Knoop C,Höckel M. Oxygenation of human tumors: evaluation of tissue oxygen distribution in breast cancers by computerized O2 tension measurements. Cancer Res 1991; 51: 3316–22.
- 24 Merritt JE,Sullivan JA,Drew L,Khan A,Wilson K,Mulqueen M,Harris W,Bradshaw D,Hill CH,Rumsby M,Warr R. The bisindolylmaleimide protein kinase C inhibitor. Ro 32-2241, reverses multidrug resistance in. KB tumour cells. Cancer Chemother Pharmacol 1999; 43: 371–8.
- 25 Barancík M,Bohácová V,Kvackajová J,Hudecová S,Krizanová O,Breier A. SB203580, a specific inhibitor of p38-MAPK pathway, is a new reversal agent of P-glycoprotein-mediated multidrug resistance. Eur J Pharm Sci 2001; 14: 29–36.
- 26 Katayama K,Yoshioka S,Tsukahara S,Mitsuhashi J,Sugimoto Y. Inhibition of the mitogen-activated protein kinase pathway results in the down-regulation of P-glycoprotein. Mol Cancer Ther 2007; 6: 2092–102.
- 27 Lin JC,Chang SY,Hsieh DS,Lee CF,Yu DS. Modulation of mitogen-activated protein kinase cascades by differentiation-1 protein: acquired drug resistance of hormone independent prostate cancer cells. J Urol 2005; 174: 2022–6.
- 28 Haworth RS,Dashnyam S,Avkiran M. Ras triggers acidosis-induced activation of the extracellular-signal-regulated kinase pathway in cardiac myocytes. Biochem J 2006; 399: 493–501.
- 29 Stathopoulou K,Gaitanaki C,Beis I. Extracellular pH changes activate the p38-MAPK signalling pathway in the amphibian heart. J Exp Biol 2006; 209: 1344–54.
- 30 Papatsoris AG,Papavassiliou AG. Prostate cancer: horizons in the development of novel anti-cancer strategies. Curr Med Chem Anticancer Agents 2001; 1: 47–70.
- 31 Cai C,Omwancha J,Hsieh CL,Shemshedini L. Androgen induces expression of the multidrug resistance protein gene MRP4 in prostate cancer cells. Prostate Cancer Prostatic Dis 2007; 10: 39–45.
- 32 van Brussel JP,Jan Van SG,Van KC,Bogdanowicz JF,Van Der Kwast TH,Schroder FH,Mickisch GH. Expression of multidrug resistance related proteins and proliferative activity is increased in advanced clinical prostate cancer. J Urol 2001; 165: 130–5.
- 33 van Brussel JP,Mickisch GH. Multidrug resistance in prostate cancer. Onkologie 2003; 26: 175–81.
- 34
Kunikane H,Zalupski MM,Ramachandran C,KuKuruga MA,Lucas D,Ryan JR,Krishan A.
Flow cytometric analysis of p-glycoprotein expression and drug efflux in human soft tissue and bone sarcomas.
Cytometry
1997;
30:
197–203.
10.1002/(SICI)1097-0320(19970815)30:4<197::AID-CYTO7>3.0.CO;2-H CAS PubMed Web of Science® Google Scholar
- 35 van de Water FM,Boleij JM,Peters JG,Russel FG,Masereeuw R. Characterization of P-glycoprotein and multidrug resistance proteins in rat kidney and intestinal cell lines. Eur J Pharm Sci 2007; 30: 36–44.
- 36 Ziemann C,Riecke A,Rüdell G,Oetjen E,Steinfelder HJ,Lass C,Kahl GF,Hirsch-Ernst KI. The role of prostaglandin E receptor- dependent signaling via cAMP in Mdr1b gene activation in primary rat hepatocyte cultures. J Pharmacol Exp Ther 2006; 317: 378–86.
- 37 Chambers TC,Pohl J,Raynor RL,Kuo JF. Identification of specific sites in human P-glycoprotein phosphorylated by protein kinase C. J Biol Chem 1993; 268: 4592–5.
- 38 Glavy JS,Horwitz SB,Orr GA. Identification of the in vivo phosphorylation sites for acidic-directed kinases in murine mdr1b P-glycoprotein. J Biol Chem 1997; 272: 5909–14.
- 39 Price DT,Della Rocca G,Guo C,Ballo MS,Schwinn DA,Luttrell LM. Activation of extracellular signal-regulated kinase in human prostate cancer. J Urol 1999; 162: 1537–42.
- 40 Ricote M,García-Tuñón I,Bethencourt F,Fraile B,Onsurbe P,Paniagua R,Royuela M. The p38 transduction pathway in prostatic neoplasia. J Pathol 2006; 208: 401–7.
- 41 McCubrey JA,Steelman LS,Chappell WH,Abrams SL,Wong EW,Chang F,Lehmann B,Terrian DM,Milella M,Tafuri A,Stivala F,Libra M, et al. Roles of the Raf/MEK/ERK pathway in cell growth, malignant transformation and drug resistance. Biochim Biophys Acta 2007; 1773: 1263–84.
- 42 Seuwen K,Ludwig MG,Wolf RM. Receptors for protons or lipid messengers or both? J Recept Signal Transduct Res 2006; 26: 599–610.
- 43 Bektas M,Barak LS,Jolly PS,Liu H,Lynch KR,Lacana E,Suhr KB,Milstien S,Spiegel S. The G protein-coupled receptor GPR4 suppresses ERK activation in a ligand-independent manner. Biochemistry 2003; 42: 12181–91.
- 44 Tomura H,Wang JQ,Komachi M,Damirin A,Mogi C,Tobo M,Kon J,Misawa N,Sato K,Okajima F. Prostaglandin I2 production and cAMP accumulation in response to acidic extracellular pH through OGR1 in human aortic smooth muscle cells. J Biol Chem 2005; 280: 34458–64.
- 45 Gekle M,Drumm K,Mildenberger S,Freudinger R,Gassner B,Silbernagl S. Inhibition of Na+-H+ exchange impairs receptor-mediated albumin endocytosis in renal proximal tubule-derived epithelial cells from opossum. J Physiol 1999; 520 Pt 3: 709–21.
- 46 Bourke VA,Zhao D,Gilio J,Chang CH,Jiang L,Hahn EW,Mason RP. Correlation of radiation response with tumor oxygenation in the Dunning prostate R3327-AT1 tumor. Int J Radiat Oncol Biol Phys 2007; 67: 1179–86.
- 47 Thews O,Kelleher DK,Lecher B,Vaupel P. Effect of cell line and differentiation on the oxygenation status of experimental sarcomas. Adv Exp Med Biol 1997; 428: 123–8.
- 48 Yeh KA,Biade S,Lanciano RM,Brown DQ,Fenning MC,Babb JS,Hanks GE,Chapman JD. Polarographic needle electrode measurements of oxygen in rat prostate carcinomas: accuracy and reproducibility. Int J Radiat Oncol Biol Phys 1995; 33: 111–18.
- 49 Chaplin DJ,Horsman MR,Trotter MJ,Siemann DW. Therapeutic significance of microenvironmental factors. In: M Molls, P Vaupel, eds. Blood perfusion and microenvironment of human tumors. Berlin: Springer, 1998. 131–43.
- 50 Di Marco A,Casazza AM,Dasdia T,Necco A,Pratesi G,Rivolta P,Velcich A,Zaccara A,Zunino F. Changes of activity of daunorubicin, adriamycin and stereoisomers following the introduction or removal of hydroxyl groups in the amino sugar moiety. Chem Biol Interact 1977; 19: 291–302.
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