title: A critical dose of doxorubicin is required to alter the gene expression profiles in MCF-7 cells acquiring multidrug resistance.
abstract: Cellular mechanisms of multidrug resistance (MDR) are related to ABC transporters, apoptosis, antioxidation, drug metabolism, DNA repair and cell proliferation. It remains unclear whether the process of resistance development is programmable. We aimed to study gene expression profiling circumstances in MCF-7 during MDR development. Eleven MCF-7 sublines with incremental doxorubicin resistance were established as a valued tool to study resistance progression. MDR marker P-gp was overexpressed only in cells termed MCF-7/ADR-1024 under the selection dose approaching 1024 nM. MCF-7/ADR-1024 and authentic MCF-7/ADR shared common features in cell morphology and DNA ploidy status. MCF-7/ADR-1024 and authentic MCF-7/ADR down regulated repair genes BRCA1/2 and wild type p53, apoptosis-related gene Bcl-2 and epithelial-mesenchymal transition (EMT) epithelial marker gene E-cadherin. While detoxifying enzymes glutathione-S transferase-π and protein kinase C-α were up-regulated. The genes involving in EMT mesenchymal formation were also overexpressed, including N-cadherin, vimentin and the E-cadherin transcription reppressors Slug, Twist and ZEB1/2. PI3K/AKT inhibitor wortmannin suppressed expression of Slug, Twist and mdr1. Mutant p53 with a deletion at codons 127-133 markedly appeared in MCF-7/ADR-1024 and authentic MCF-7/ADR as well. In addition, MCF-7/ADR-1024 cells exerted CSC-like cell surface marker CD44 high/CD24 low and form mammospheres. Overall, results suggest that resistance marker P-gp arises owing to turn on/off or mutation of the genes involved in DNA repair, apoptosis, detoxifying enzymes, EMT and ABC transporters at a turning point (1.024 μM doxorubicin challenge). Behind this point, no obvious alterations were found in most tested genes. Selection for CSC-like cells under this dose may importantly attribute to propagation of the population presenting invasive properties and drug resistance. We thereby suggest two models in the induction of drug resistance. Model 1: Selection for CSC-like cells. Model 2: Mutations for gain-of resistance. Either model 1 or model 2 requires doxorubicin dose approaching 1 μM to alter gene regulation.
authors: Graduate Institute of Pharmaceutical Sciences, School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors: Shang-Hsun
authors: SH
authors: Tsou
authors: Department of Obstetrics and Gynecology, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors: Tzer-Ming
authors: TM
authors: Chen
authors: Graduate Institute of Pharmaceutical Sciences, School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors: Hui-Ting
authors: HT
authors: Hsiao
authors: Graduate Institute of Pharmaceutical Sciences, School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan; Graduate Institute of Clinical Pharmacy, School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Taiwan.
authors: Yen-Hui
authors: YH
authors: Chen
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001943:Breast Neoplasms / Q000235:genetics*; D048430:Cell Shape / Q000187:drug effects; D004260:DNA Repair / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects; D004317:Doxorubicin / Q000494:pharmacology*; D018432:Drug Resistance, Multiple / Q000187:drug effects / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000187:drug effects / Q000235:genetics*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects / Q000235:genetics; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D052138:Genes, Neoplasm; D006801:Humans; D020128:Inhibitory Concentration 50; D061986:MCF-7 Cells; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D014411:Neoplastic Stem Cells / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D009363:Neoplasm Proteins; D016159:Tumor Suppressor Protein p53; D004317:Doxorubicin
references: Med Princ Pract. 2005;14 Suppl 1:35-48
references: 16103712
references: Science. 1990 Dec 14;250(4987):1576-80
references: 2274789
references: Cancer Lett. 2012 Dec 28;325(2):165-74
references: 22771536
references: J Biol Chem. 2001 Oct 19;276(42):39359-67
references: 11483599
references: Antioxid Redox Signal. 2009 Jan;11(1):99-133
references: 18699730
references: Oncogene. 1997 Jan 30;14(4):499-506
references: 9053847
references: Cancer Sci. 2011 Apr;102(4):663-9
references: 21205087
references: Biochemistry (Mosc). 2008 May;73(5):592-604
references: 18605983
references: Exp Oncol. 2009 Sep;31(3):140-3
references: 19783968
references: Neoplasma. 2012;59(3):248-56
references: 22329846
references: Toxicol Sci. 2001 Jun;61(2):218-23
references: 11353130
references: IUBMB Life. 2007 Dec;59(12):752-7
references: 18085475
references: Int J Cancer. 2005 Oct 20;117(1):21-31
references: 15880572
references: Int J Cancer. 2009 Feb 15;124(4):806-15
references: 19035454
references: Mol Cell Biochem. 2002 May-Jun;234-235(1-2):249-63
references: 12162441
references: Biol Pharm Bull. 2001 May;24(5):474-9
references: 11379763
references: J Natl Cancer Inst. 2010 Nov 3;102(21):1637-52
references: 20935265
references: Cell Death Dis. 2011;2:e179
references: 21734725
references: Cancer Res. 1993 Oct 1;53(19):4701-14
references: 8402648
references: Cell Biochem Biophys. 2009;55(2):95-105
references: 19593673
references: Crit Rev Oncog. 1994;5(1):23-57
references: 7948107
references: Int J Cancer. 1996 Sep 4;67(5):608-14
references: 8782646
references: Anticancer Res. 2007 May-Jun;27(3A):1267-72
references: 17593618
references: Mini Rev Med Chem. 2010 May;10(5):359-71
references: 20370703
references: Toxicol Sci. 2002 Dec;70(2):183-92
references: 12441363
references: Oncogene. 2003 Oct 20;22(47):7486-95
references: 14576853
references: Biochem Biophys Res Commun. 2012 Mar 23;419(4):728-34
references: 22387537
references: Anticancer Drugs. 2009 Jun;20(5):382-8
references: 19318911
references: J Natl Cancer Inst. 2002 Nov 6;94(21):1652-4; author reply 1654
references: 12419794
references: Toxicol Appl Pharmacol. 2010 Apr 1;244(1):66-76
references: 19732782
references: Asian Pac J Cancer Prev. 2012;13(8):3631-6
references: 23098446
references: Clin Transl Oncol. 2012 Oct;14(10):715-20
references: 22911547
references: Oncogene. 2004 Sep 20;23(43):7274-82
references: 15378087
references: Mol Cancer. 2010;9:145
references: 20540746
references: Blood. 2003 Aug 15;102(4):1466-73
references: 12689934
references: Biomed Pharmacother. 2011 Feb;65(1):40-5
references: 21177063
references: BMC Cancer. 2011;11:418
references: 21957977
references: Cell Growth Differ. 1995 Nov;6(11):1395-403
references: 8562478
references: J Clin Invest. 2001 Mar;107(5):643-6
references: 11238567
references: PLoS One. 2008;3(8):e2888
references: 18682804
references: APMIS. 2005 Nov-Dec;113(11-12):922-9
references: 16480458
references: J Biol Chem. 2009 Jan 30;284(5):2657-71
references: 19047049
references: Free Radic Biol Med. 2009 Dec 1;47(11):1619-31
references: 19751820
references: Biochim Biophys Acta. 1976 Nov 11;455(1):152-62
references: 990323
references: Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):5878-82
references: 8016082
references: J Biol Chem. 2001 Jun 15;276(24):20999-1003
references: 11279197
references: Proc Natl Acad Sci U S A. 2003 Apr 1;100(7):3983-8
references: 12629218
references: Prog Clin Biol Res. 1992;376:331-55
references: 1528927
references: Biochem Biophys Res Commun. 2007 Jun 22;358(1):79-84
references: 17482571
references: Radiat Res. 1995 Jun;142(3):256-62
references: 7761574
references: Oncogene. 2003 Oct 20;22(47):7496-511
references: 14576854
references: J Thorac Oncol. 2012 Dec;7(12):1880-90
references: 23154562
references: Nat Rev Cancer. 2002 Jan;2(1):48-58
references: 11902585
references: BMC Cancer. 2003 Apr 24;3:13
references: 12713671
references: Cancers (Basel). 2011 Jun;3(2):1639-71
references: 22348197
references: Adv Exp Med Biol. 2013;779:35-54
references: 23288634