Journal Mobile Options
Table of Contents
Vol. 9, No. 1-2, 2009
Issue release date: April 2009
Pancreatology 2009;9:136–144

ATP-Binding Cassette C Transporters in Human Pancreatic Carcinoma Cell Lines

Upregulation in 5-Fluorouracil-Resistant Cells

Hagmann W. · Jesenofsky R. · Faissner R. · Guo C. · Löhr J.M.
aClinical Cooperation Unit of Molecular Gastroenterology, German Cancer Research Center, Heidelberg, Germany; bDepartment of Surgical Gastroenterology, CLINTEC, Karolinska Institute, Stockholm, Sweden

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Background: Pancreatic cancer is characterized by high resistance to chemotherapy. Such chemoresistance can be mediated by multidrug resistance proteins (MRPs), breast cancer resistance protein (BCRP), and MDR1 P-glycoprotein. However, the contribution of individual MRP isoforms to chemoresistance in pancreatic carcinoma is unclear. We studied ATP-binding cassette (ABC) transporter expression in human pancreatic carcinoma cell lines as compared to primary pancreatic duct cells, and analyzed the MRP expression profile in 5-fluorouracil-resistant cells. Methods: Transporter expression was analyzed by quantitative and qualitative RT-PCR, by immunoblot, and chemoresistance by cytotoxicity assay. Results: Primary pancreatic duct cells expressed MRP1, MRP3, MRP4, and MRP5, but not MRP2 mRNA. The established carcinoma cell lines expressed MRP1, MRP4, and MRP5, most of them also MRP2, MRP3, MRP7, and BCRP, but none contained detectable amounts of MRP6, MRP8, or MRP9 mRNA. Immunoblot analyses demonstrated presence of MRP1, MRP4, and MRP5 protein in all, but MRP3 and BCRP protein only in some of these cells. Compared to parental Capan-1 cells, Capan-1 cells with acquired chemoresistance towards 5-fluorouracil showed an upregulated mRNA and protein expression of MRP3, MRP4, and MRP5. In addition, silencing of MRP5 by RNA interference resulted in enhanced sensitivity of parental Capan-1 cells towards 5-fluorouracil cytotoxicity. Conclusion: MRP3, MRP4, and MRP5 are upregulated in 5-fluorouracil-resistant cells, and MRP5 contributes to 5-FU resistance in pancreatic carcinoma cells.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Jemal A, Siegel R, Ward E, Murray T, Xu J, Smigal C, Thun MJ: Cancer statistics, 2006. CA Cancer J Clin 2006;56:106–130.
  2. Löhr JM: Medical treatment of pancreatic cancer. Expert Rev Anticancer Ther 2007;7:533–544.
  3. Löhr M: Is it possible to survive pancreatic cancer? Nat Clin Pract Gastroenterol Hepatol 2006;3:236–237.
  4. Erlichman C, Fine S, Wong A, Elhakim T: A randomized trial of fluorouracil and folinic acid in patients with metastatic colorectal carcinoma. J Clin Oncol 1988;6:469–475.
  5. Lokich JJ, Ahlgren JD, Gullo JJ, Philips JA, Fryer JG: A prospective randomized comparison of continuous infusion fluorouracil with a conventional bolus schedule in metastatic colorectal carcinoma: a mid-Atlantic oncology program study. J Clin Oncol 1989;7:425–432.
  6. Miyashita T, Reed JC: Bcl-2 oncoprotein blocks chemotherapy-induced apoptosis in a human leukemia cell line. Blood 1993;81:151–157.
  7. Korsmeyer SJ: Bcl-2 initiates a new category of oncogenes: regulators of cell death. Blood 1992;80:879–886.
  8. Borst P, Evers R, Kool M, Wijnholds J: A family of drug transporters: the multidrug resistance-associated proteins. J Natl Cancer Inst 2000;92:1295–1302.
  9. Kruh GD, Belinsky MG: The MRP family of drug efflux pumps. Oncogene 2003;22:7537–7552.
  10. Borst P, de Wolf C, van de Wetering K: Multidrug resistance-associated proteins 3, 4, and 5. Pflügers Arch 2007;453:661–673.
  11. König J, Nies AT, Cui Y, Leier I, Keppler D: Conjugate export pumps of the multidrug resistance protein (MRP) family: localization, substrate specificity, and MRP2-mediated drug resistance. Biochim Biophys Acta 1999;1461:377–394.
  12. Szakacs G, Paterson JK, Ludwig JA, Booth-Genthe C, Gottesman MM: Targeting multidrug resistance in cancer. Nat Rev Drug Discov 2006;5:219–234.
  13. Cui Y, König J, Buchholz JK, Spring H, Leier I, Keppler D: Drug resistance and ATP-dependent conjugate transport mediated by the apical multidrug resistance protein, MRP2, permanently expressed in human and canine cells. Mol Pharmacol 1999;55:929–937.
  14. Leslie EM, Deeley RG, Cole SP: Multidrug resistance proteins: role of p-glycoprotein, MRP1, MRP2, and BCRP (ABCG2) in tissue defense. Toxicol Appl Pharmacol 2005;204:216–237.
  15. Suwa H, Ohshio G, Arao S, Imamura T, Yamaki K, Manabe T, Imamura M, Hiai H, Fukumoto M: Immunohistochemical localization of p-glycoprotein and expression of the multidrug resistance-1 gene in human pancreatic cancer: relevance to indicator of better prognosis. Jpn J Cancer Res 1996;87:641–649.
  16. Lu Z, Kleeff J, Shrikhande S, Zimmermann T, Korc M, Friess H, Büchler MW: Expression of the multidrug-resistance 1 (MDR1) gene and prognosis in human pancreatic cancer. Pancreas 2000;21:240–247.
  17. Kiuchi Y, Suzuki H, Hirohashi T, Tyson CA, Sugiyama Y: cDNA cloning and inducible expression of human multidrug resistance associated protein 3 (MRP3). FEBS Lett 1998;433:149–152.
  18. König J, Rost D, Cui Y, Keppler D: Characterization of the human multidrug resistance protein isoform MRP3 localized to the basolateral hepatocyte membrane. Hepatology 1999;29:1156–1163.
  19. Kool M, van der Linden M, de Haas M, Scheffer GL, de Vree JM, Smith AJ, Jansen G, Peters GJ, Ponne N, Scheper RJ, Elferink RP, Baas F, Borst P: MRP3, an organic anion transporter able to transport anti-cancer drugs. Proc Natl Acad Sci USA 1999;96:6914–6919.
  20. Scheffer GL, Kool M, de Haas M, de Vree JM, Pijnenborg AC, Bosman DK, Elferink RP, van der Valk P, Borst P, Scheper RJ: Tissue distribution and induction of human multidrug resistant protein 3. Lab Invest 2002;82:193–201.
  21. Schaarschmidt T, Merkord J, Adam U, Schroeder E, Kunert-Keil C, Sperker B, Drewelow B, Wacke R: Expression of multidrug resistance proteins in rat and human chronic pancreatitis. Pancreas 2004;28:45–52.
  22. Sandusky GE, Mintze KS, Pratt SE, Dantzig AH: Expression of multidrug resistance-associated protein 2 (MRP2) in normal human tissues and carcinomas using tissue microarrays. Histopathology 2002;41:65–74.
  23. König J, Hartel M, Nies AT, Martignoni ME, Guo J, Büchler MW, Friess H, Keppler D: Expression and localization of human multidrug resistance protein (ABCC) family members in pancreatic carcinoma. Int J Cancer 2005;115:359–367.
  24. Slitt AL, Cherrington NJ, Fisher CD, Negishi M, Klaassen CD: Induction of genes for metabolism and transport by trans-stilbene oxide in livers of Sprague-Dawley and Wistar-Kyoto rats. Drug Metab Dispos 2006;34:1190–1197.
  25. Jesnowski R, Müller P, Schareck W, Liebe S, Löhr M: Immortalized pancreatic duct cells in vitro and in vivo. Ann NY Acad Sci 1999;880:50–65.
  26. Sipos B, Moser S, Kalthoff H, Torok V, Löhr M, Klöppel G: A comprehensive characterization of pancreatic ductal carcinoma cell lines: towards the establishment of an in vitro research platform. Virchows Arch 2003;442:444–452.
  27. Moore PS, Sipos B, Orlandini S, Sorio C, Real FX, Lemoine NR, Gress T, Bassi C, Klöppel G, Kalthoff H, Ungefroren H, Löhr M, Scarpa A: Genetic profile of 22 pancreatic carcinoma cell lines: analysis of k-ras, p53, p16 and dpc4/smad4. Virchows Arch 2001;439:798–802.
  28. Jesnowski R, Fürst D, Ringel J, Chen Y, Schrödel A, Kleeff J, Kolb A, Schareck WD, Löhr M: Immortalization of pancreatic stellate cells as an in vitro model of pancreatic fibrosis: deactivation is induced by matrigel and n-acetylcysteine. Lab Invest 2005;85:1276–1291.
  29. Schinkel AH, Wagenaar E, van Deemter L, Mol CA, Borst P: Absence of the mdr1a p-glycoprotein in mice affects tissue distribution and pharmacokinetics of dexamethasone, digoxin, and cyclosporin A. J Clin Invest 1995;96:1698–1705.
  30. Paumi CM, Wright M, Townsend AJ, Morrow CS: Multidrug resistance protein (MRP) 1 and MRP3 attenuate cytotoxic and transactivating effects of the cyclopentenone prostaglandin, 15-deoxy-delta(12,14)prostaglandin J2 in MCF7 breast cancer cells. Biochemistry 2003;42:5429–5437.
  31. Bortfeld M, Rius M, König J, Herold-Mende C, Nies AT, Keppler D: Human multidrug resistance protein 8 (MRP8/ABCC11), an apical efflux pump for steroid sulfates, is an axonal protein of the CNS and peripheral nervous system. Neuroscience 2006;137:1247–1257.
  32. Rius M, Nies AT, Hummel-Eisenbeiss J, Jedlitschky G, Keppler D: Cotransport of reduced glutathione with bile salts by MRP4 (ABCC4) localized to the basolateral hepatocyte membrane. Hepatology 2003;38:374–384.
  33. Jedlitschky G, Burchell B, Keppler D: The multidrug resistance protein 5 functions as an ATP-dependent export pump for cyclic nucleotides. J Biol Chem 2000;275:30069–30074.
  34. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–254.
  35. Hagmann W, Nies AT, König J, Frey M, Zentgraf H, Keppler D: Purification of the human apical conjugate export pump MRP2: reconstitution and functional characterization as substrate-stimulated ATPase. Eur J Biochem 1999;265:281–289.
  36. Nishimura M, Naito S: Tissue-specific mRNA expression profiles of human ATP-binding cassette and solute carrier transporter superfamilies. Drug Metab Pharmacokinet 2005;20:452–477.
  37. Jesnowski R, Backhaus C, Ringel J, Löhr M: Ribosomal highly basic 23-kDa protein as a reliable standard for gene expression analysis. Pancreatology 2002;2:421–424.
  38. Haber PS, Keogh GW, Apte MV, Moran CS, Stewart NL, Crawford DH, Pirola RC, McCaughan GW, Ramm GA, Wilson JS: Activation of pancreatic stellate cells in human and experimental pancreatic fibrosis. Am J Pathol 1999;155:1087–1095.
  39. Yen TW, Aardal NP, Bronner MP, Thorning DR, Savard CE, Lee SP, Bell RH Jr: Myofibroblasts are responsible for the desmoplastic reaction surrounding human pancreatic carcinomas. Surgery 2002;131:129–134.
  40. Schneiderhan W, Diaz F, Fundel M, Zhou S, Siech M, Hasel C, Möller P, Gschwend JE, Seufferlein T, Gress T, Adler G, Bachem MG: Pancreatic stellate cells are an important source of MMP-2 in human pancreatic cancer and accelerate tumor progression in a murine xenograft model and CAM assay. J Cell Sci 2007;120:512–519.
  41. van Herwaarden AE, Schinkel AH: The function of breast cancer resistance protein in epithelial barriers, stem cells and milk secretion of drugs and xenotoxins. Trends Pharmacol Sci 2006;27:10–16.
  42. Jedlitschky G, Leier I, Buchholz U, Hummel-Eisenbeiss J, Burchell B, Keppler D: ATP-dependent transport of bilirubin glucuronides by the multidrug resistance protein MRP1 and its hepatocyte canalicular isoform MRP2. Biochem J 1997;327(part 1):305–310.
  43. Bailey-Dell KJ, Hassel B, Doyle LA, Ross DD: Promoter characterization and genomic organization of the human breast cancer resistance protein (ATP-binding cassette transporter G2) gene. Biochim Biophys Acta 2001;1520:234–241.
  44. Hagmann W, Schubert J, König J, Keppler D: Reconstitution of transport-active multidrug resistance protein 2 (MRP2; ABCC2) in proteoliposomes. Biol Chem 2002;383:1001–1009.
  45. Pratt S, Shepard RL, Kandasamy RA, Johnston PA, Perry W 3rd, Dantzig AH: The multidrug resistance protein 5 (ABCC5) confers resistance to 5-fluorouracil and transports its monophosphorylated metabolites. Mol Cancer Ther 2005;4:855–863.
  46. Oguri T, Bessho Y, Achiwa H, Ozasa H, Maeno K, Maeda H, Sato S, Ueda R: MRP8/ABCC11 directly confers resistance to 5-fluorouracil. Mol Cancer Ther 2007;6:122–127.
  47. Monti P, Marchesi F, Reni M, Mercalli A, Sordi V, Zerbi A, Balzano G, Di Carlo V, Allavena P, Piemonti L: A comprehensive in vitro characterization of pancreatic ductal carcinoma cell line biological behavior and its correlation with the structural and genetic profile. Virchows Arch 2004;445:236–247.
  48. Shi X, Liu S, Kleeff J, Friess H, Büchler MW: Acquired resistance of pancreatic cancer cells towards 5-fluorouracil and gemcitabine is associated with altered expression of apoptosis-regulating genes. Oncology 2002;62:354–362.
  49. Young LC, Campling BG, Cole SP, Deeley RG, Gerlach JH: Multidrug resistance proteins MRP3, MRP1, and MRP2 in lung cancer: correlation of protein levels with drug response and messenger RNA levels. Clin Cancer Res 2001;7:1798–1804.
  50. Rius M, Hummel-Eisenbeiss J, Hofmann AF, Keppler D: Substrate specificity of human ABCC4 (MRP4)-mediated cotransport of bile acids and reduced glutathione. Am J Physiol Gastrointest Liver Physiol 2006;290:G640–G649.
  51. Guo Y, Kotova E, Chen ZS, Lee K, Hopper-Borge E, Belinsky MG, Kruh GD: MRP8, ATP-binding cassette C11 (ABCC11), is a cyclic nucleotide efflux pump and a resistance factor for fluoropyrimidines 2′,3′-dideoxycytidine and 9′-(2′-phosphonylmethoxyethyl)adenine. J Biol Chem 2003;278:29509–29514.
  52. Lee JH, Park JH, Jung Y, Kim JH, Jong HS, Kim TY, Bang YJ: Histone deacetylase inhibitor enhances 5-fluorouracil cytotoxicity by down-regulating thymidylate synthase in human cancer cells. Mol Cancer Ther 2006;5:3085–3095.
  53. Wang W, McLeod HL, Cassidy J, Collie-Duguid ES: Mechanisms of acquired chemoresistance to 5-fluorouracil and tomudex: thymidylate synthase dependent and independent networks. Cancer Chemother Pharmacol 2007;59:839–845.
  54. Ooyama A, Okayama Y, Takechi T, Sugimoto Y, Oka T, Fukushima M: Genome-wide screening of loci associated with drug resistance to 5-fluorouracil-based drugs. Cancer Sci 2007;98:577–583.
  55. Shah AN, Summy JM, Zhang J, Park SI, Parikh NU, Gallick GE: Development and characterization of gemcitabine-resistant pancreatic tumor cells. Ann Surg Oncol 2007;14:3629–3637.
  56. Arlt A, Gehrz A, Müerköster S, Vorndamm J, Kruse ML, Fölsch UR, Schäfer H: Role of NF-kB and Akt/PI3K in the resistance of pancreatic carcinoma cell lines against gemcitabine-induced cell death. Oncogene 2003;22:3243–3251.
  57. Kuo MT, Liu Z, Wei Y, Lin-Lee YC, Tatebe S, Mills GB, Unate H: Induction of human MDR1 gene expression by 2-acetylaminofluorene is mediated by effectors of the phosphoinositide 3-kinase pathway that activate NF-kB signaling. Oncogene 2002;21:1945–1954.
  58. Sims-Mourtada J, Izzo JG, Ajani J, Chao KS: Sonic Hedgehog promotes multiple drug resistance by regulation of drug transport. Oncogene 2007;26:5674–5679.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50