title: Interdependence of gemcitabine treatment, transporter expression, and resistance in human pancreatic carcinoma cells.
abstract: Gemcitabine is widely used as first-line chemotherapeutic drug in the treatment of pancreatic cancer. Our previous experimental chemotherapy studies have shown that treatment of human pancreatic carcinoma cells with 5-fluorouracil (5-FU) alters the cellular transporter expression profile and that modulation of the expression of multidrug resistance protein 5 (MRP5; ABCC5) influences the chemoresistance of these tumor cells. Here, we studied the influence of acute and chronic gemcitabine treatment on the expression of relevant uptake and export transporters in pancreatic carcinoma cells by reverse transcription-polymerase chain reaction (RT-PCR), quantitative RT-PCR, and immunoblot analyses. The specific role of MRP5 in cellular gemcitabine sensitivity was studied by cytotoxicity assays using MRP5-overexpressing and MRP5-silenced cells. Exposure to gemcitabine (12 nM for 3 days) did not alter the messenger RNA (mRNA) expression of MRP1, MRP3, MRP5, and equilibrative nucleoside transporter 1 (ENT1), whereas high dosages of the drug (20 microM for 1 hour) elicited up-regulation of these transporters in most cell lines studied. In cells with acquired gemcitabine resistance (up to 160 nM gemcitabine), the mRNA or protein expression of the gemcitabine transporters MRP5 and ENT1 was upregulated in several cell lines. Combined treatment with 5-FU and gemcitabine caused a 5- to 40-fold increase in MRP5 and ENT1 expressions. Cytotoxicity assays using either MRP5-overexpressing (HEK and PANC-1) or MRP5-silenced (PANC1/shMRP5) cells indicated that MRP5 contributes to gemcitabine resistance. Thus, our novel data not only on drug-induced alterations of transporter expression relevant for gemcitabine uptake and export but also on the link between gemcitabine sensitivity and MRP5 expression may lead to improved strategies of future chemotherapy regimens using gemcitabine in pancreatic carcinoma patients.
authors.affiliation: Clinical Cooperation Unit of Molecular Gastroenterology, German Cancer Research Center, Heidelberg, Germany. w.hagmann@dkfz.de
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hagmann
authors.forename: Ralf
authors.initials: R
authors.lastname: Jesnowski
authors.forename: Johannes Matthias
authors.initials: JM
authors.lastname: Löhr
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000009:adverse effects / Q000494:pharmacology / Q000627:therapeutic use; D002277:Carcinoma / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D003841:Deoxycytidine / Q000009:adverse effects / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm / Q000187:drug effects* / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D010190:Pancreatic Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D013997:Time Factors; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics; D000093542:Gemcitabine
chemical_list: D000964:Antimetabolites, Antineoplastic; D002352:Carrier Proteins; D003841:Deoxycytidine; D000093542:Gemcitabine
references.citation: J Biol Chem. 2001 Dec 7;276(49):46400-7
references.pmid: 11581266
references.citation: Clin Cancer Res. 2009 Apr 15;15(8):2913-9
references.pmid: 19318496
references.citation: Cancer Res. 2000 Nov 1;60(21):6075-9
references.pmid: 11085530
references.citation: Lab Invest. 2002 Feb;82(2):193-201
references.pmid: 11850532
references.citation: Cancer Res. 2010 Feb 1;70(3):1130-40
references.pmid: 20103639
references.citation: Proc Natl Acad Sci U S A. 1999 Jun 8;96(12):6914-9
references.pmid: 10359813
references.citation: Oncology. 2002;62(4):354-62
references.pmid: 12138244
references.citation: Lancet. 2004 Mar 27;363(9414):1049-57
references.pmid: 15051286
references.citation: Clin Cancer Res. 2008 Mar 1;14(5):1470-7
references.pmid: 18316571
references.citation: Oncol Rep. 2007 May;17(5):1201-5
references.pmid: 17390066
references.citation: Mol Pharmacol. 2003 May;63(5):1094-103
references.pmid: 12695538
references.citation: Biochem Pharmacol. 2008 Aug 1;76(3):322-9
references.pmid: 18589402
references.citation: Dig Dis Sci. 2009 Jan;54(1):89-96
references.pmid: 18594980
references.citation: Gastroenterology. 2009 Jan;136(1):187-95
references.pmid: 18992248
references.citation: Eur J Pharm Sci. 2006 Apr;27(5):524-32
references.pmid: 16337112
references.citation: Cancer Res. 1998 Oct 1;58(19):4349-57
references.pmid: 9766663
references.citation: Pancreas. 2009 Oct;38(7):e190-9
references.pmid: 19730150
references.citation: Clin Cancer Res. 1997 Nov;3(11):2025-32
references.pmid: 9815593
references.citation: Clin Cancer Res. 2004 May 1;10(9):2936-43
references.pmid: 15131028
references.citation: Expert Rev Anticancer Ther. 2002 Aug;2(4):426-36
references.pmid: 12647986
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Br J Cancer. 2007 Feb 12;96(3):457-63
references.pmid: 17224927
references.citation: CA Cancer J Clin. 2009 Jul-Aug;59(4):225-49
references.pmid: 19474385
references.citation: Mol Cancer Ther. 2005 May;4(5):855-63
references.pmid: 15897250
references.citation: Cancer Res. 2001 Jan 15;61(2):550-5
references.pmid: 11212248
references.citation: Pflugers Arch. 2007 Feb;453(5):661-73
references.pmid: 16586096
references.citation: Semin Oncol. 2007 Aug;34(4):347-53
references.pmid: 17674963
references.citation: J Natl Cancer Inst. 1999 Nov 3;91(21):1876-81
references.pmid: 10547395
references.citation: J Biol Chem. 2001 Jan 26;276(4):2914-27
references.pmid: 11032837
references.citation: Int J Cancer. 2005 Jun 20;115(3):359-67
references.pmid: 15688370
references.citation: Expert Rev Anticancer Ther. 2003 Oct;3(5):729-39
references.pmid: 14599095
references.citation: Biochem Pharmacol. 2010 Feb 15;79(4):587-95
references.pmid: 19788890
references.citation: Clin Cancer Res. 2009 Jun 1;15(11):3705-15
references.pmid: 19458058
references.citation: FEBS Lett. 1998 Aug 14;433(1-2):149-52
references.pmid: 9738950
references.citation: Int J Cancer. 2009 Nov 15;125(10):2323-31
references.pmid: 19598259
references.citation: Clin Cancer Res. 2003 Oct 15;9(13):5000-8
references.pmid: 14581375
references.citation: Cancer Metastasis Rev. 2007 Mar;26(1):85-110
references.pmid: 17345146
references.citation: Mol Cancer Ther. 2006 Jul;5(7):1800-6
references.pmid: 16891466
references.citation: Scand J Gastroenterol. 2009;44(7):782-6
references.pmid: 19214867
references.citation: Br J Cancer. 1995 Oct;72(4):939-42
references.pmid: 7547244
references.citation: Mol Pharmacol. 1999 May;55(5):929-37
references.pmid: 10220572
references.citation: Cancer Res. 2006 Apr 1;66(7):3928-35
references.pmid: 16585222
references.citation: Stem Cells. 2006 Mar;24(3):506-13
references.pmid: 16239320
references.citation: Clin Cancer Res. 2004 Apr 1;10(7):2307-18
references.pmid: 15073106
references.citation: Clin Cancer Res. 2008 Dec 15;14(24):8143-51
references.pmid: 19088029
references.citation: Mol Cancer Ther. 2009 Apr;8(4):809-20
references.pmid: 19372554
references.citation: Pancreatology. 2009;9(1-2):136-44
references.pmid: 19077464
references.citation: Gastrointest Cancer Res. 2009 Mar;3(2 Suppl):S11-5
references.pmid: 19461915
references.citation: Pancreas. 2006 Aug;33(2):142-7
references.pmid: 16868479
references.citation: Biochim Biophys Acta. 1999 Dec 6;1461(2):377-94
references.pmid: 10581368
references.citation: Eur J Biochem. 1999 Oct 1;265(1):281-9
references.pmid: 10491184
references.citation: BMC Cancer. 2008 Mar 28;8:82
references.pmid: 18373843
references.citation: Nat Clin Pract Gastroenterol Hepatol. 2006 May;3(5):236-7
references.pmid: 16672986
references.citation: Cancer Chemother Pharmacol. 1991;27(4):258-62
references.pmid: 1998982
references.citation: J Clin Oncol. 1997 Jun;15(6):2403-13
references.pmid: 9196156
references.citation: Br J Cancer. 1996 Jan;73(1):101-5
references.pmid: 8554969
references.citation: Clin Cancer Res. 2004 Oct 15;10(20):6956-61
references.pmid: 15501974
references.citation: Hepatology. 1999 Apr;29(4):1156-63
references.pmid: 10094960
references.citation: J Biol Chem. 2000 Sep 29;275(39):30069-74
references.pmid: 10893247