title: Pluripotency markers are differentially induced by MEK inhibition in thyroid and melanoma BRAFV600E cell lines.
abstract: Oncogenic mutations in BRAF are common in melanoma and thyroid carcinoma and drive constitutive activation of the MAPK pathway. Molecularly targeted therapies of this pathway improves survival compared to chemotherapy; however, responses tend to be short-lived as resistance invariably occursCell line models of melanoma and thyroid carcinoma, +/- BRAF(V600E) activating mutation, were treated with the MEK inhibitor PD0325901. Treated and naive samples were assayed for expression of key members of the MAPK pathway. Global microRNA expression profiling of naive and resistant cells was performed via next generation sequencingand indicated pluripotency pathways in resistance. Parental cell lines were progressed to holoclones to confirm the miRNA stemness profileMembers of the MIR302/373/374/520 family of embryonic stem cell specific cell cycle regulating (ESCC) microRNAs were identified as differentially expressed between resistant BRAF(V600E) melanoma and thyroid cell lines. Upregulated expression of gene and protein stemness markers, upregulated expression of MAPK pathway genes and downregulation of the ESCC MIR302 cluster in BRAF(V600E) melanoma indicated an increased stem-like phenotype in resistant BRAF(V600E) melanoma. Conversely, downregulated expression of gene and protein stemness markers, downregulated expression of MAPK pathway genes, upregulation of the ESCC MIR520 cluster, reeexpression of cell surface receptors, and induced differentiation-associated morphology in resistant BRAF(V600E) indicate a differentiated phenotype associated with MEK inhibitor resistance in BRAF(V600E) thyroid cellsThe differential patterns of resistance observed between BRAF(V600E) melanoma and thyroid cell lines may reflect tissue type or de novo differentiation, but could have significant impact on the response of primary and metastatic cells to MEK inhibitor treatment. This study provides a basis for the investigation of the cellular differentiation/self-renewal access and its role in resistance to MEK inhibition.
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: Emma R
authors.initials: ER
authors.lastname: Dorris
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: Gordon
authors.initials: G
authors.lastname: Blackshields
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: Gary
authors.initials: G
authors.lastname: Sommerville
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Alhashemi
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: Andrew
authors.initials: A
authors.lastname: Dias
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: Victoria
authors.initials: V
authors.lastname: McEneaney
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: Paul
authors.initials: P
authors.lastname: Smyth
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: John J
authors.initials: JJ
authors.lastname: O'Leary
authors.affiliation: a Department of Histopathology , Sir Patrick Dun Research Lab, Trinity College Dublin , Dublin , Ireland.
authors.forename: Orla
authors.initials: O
authors.lastname: Sheils
mesh_terms: D002454:Cell Differentiation; D045744:Cell Line, Tumor; D006801:Humans; D008545:Melanoma / Q000188:drug therapy*; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism*; D039904:Pluripotent Stem Cells; D013964:Thyroid Neoplasms / Q000188:drug therapy*
chemical_list: D020929:Mitogen-Activated Protein Kinase Kinases
keywords: BRAFV600E; MEK inhibition; cancer; cancer stem cell; microrna; pluripotency; resistance
references.citation: Biochem Biophys Res Commun. 2011 Jan 28;404(4):958-62
references.pmid: 21185263
references.citation: Nature. 2011 Nov 23;480(7377):387-90
references.pmid: 22113612
references.citation: FEBS Lett. 2005 Oct 31;579(26):5911-22
references.pmid: 16111679
references.citation: Semin Oncol. 2012 Apr;39(2):204-14
references.pmid: 22484192
references.citation: Nat Biotechnol. 2009 May;27(5):459-61
references.pmid: 19363475
references.citation: Nucleic Acids Res. 2011 Feb;39(3):1054-65
references.pmid: 20870751
references.citation: J Chin Med Assoc. 2010 Mar;73(3):113-28
references.pmid: 20230995
references.citation: Genes Cancer. 2011 Mar;2(3):195-209
references.pmid: 21779493
references.citation: Nature. 2004 Nov 18;432(7015):294-7
references.pmid: 15549090
references.citation: Mamm Genome. 2007 May;18(5):316-27
references.pmid: 17610011
references.citation: N Engl J Med. 2012 Jul 12;367 (2):107-14
references.pmid: 22663011
references.citation: Nature. 2010 Dec 16;468(7326):968-72
references.pmid: 21107320
references.citation: J Biol Chem. 2012 Sep 21;287(39):32800-24
references.pmid: 22847005
references.citation: J Clin Oncol. 2008 May 1;26(13):2139-46
references.pmid: 18390968
references.citation: Cancer Res. 2008 Jun 15;68(12):4853-61
references.pmid: 18559533
references.citation: Cell. 2011 Apr 1;145(1):30-8
references.pmid: 21458666
references.citation: Cell Death Differ. 2012 Feb;19(2):232-44
references.pmid: 21720384
references.citation: Nat Rev Cancer. 2005 Apr;5(4):275-84
references.pmid: 15803154
references.citation: PLoS One. 2011;6(12 ):e28688
references.pmid: 22205962
references.citation: Cold Spring Harb Symp Quant Biol. 2008;73:171-4
references.pmid: 19150965
references.citation: Genome Biol. 2010;11(10):R106
references.pmid: 20979621
references.citation: Cancer Cell. 2015 Jan 12;27(1):85-96
references.pmid: 25500121
references.citation: Nature. 2006 Jan 19;439(7074):358-62
references.pmid: 16273091
references.citation: Nucleic Acids Res. 2006 Jan 24;34(2):e9
references.pmid: 16434699
references.citation: Clin Cancer Res. 2012 Jan 15;18(2):555-67
references.pmid: 22048237
references.citation: Genome Res. 2008 Apr;18(4):610-21
references.pmid: 18285502
references.citation: Nat Biotechnol. 2011 May;29(5):443-8
references.pmid: 21490602
references.citation: J Carcinog. 2004 Mar 26;3(1):6
references.pmid: 15046639
references.citation: Cancer Res. 2010 Nov 15;70(22):9473-82
references.pmid: 21062975
references.citation: J Immunol. 2012 Jun 15;188(12):6357-70
references.pmid: 22581862
references.citation: N Engl J Med. 2014 Nov 13;371(20):1877-88
references.pmid: 25265492
references.citation: Mol Cancer Ther. 2010 Jul;9(7):1968-76
references.pmid: 20587665
references.citation: Prog Clin Biol Res. 1980;48:291-312
references.pmid: 7193873
references.citation: FEBS Lett. 2009 Jun 18;583(12):1817-24
references.pmid: 19433088
references.citation: J Cell Mol Med. 2007 Nov-Dec;11(6):1251-62
references.pmid: 18205699
references.citation: Curr Biol. 2011 Jan 11;21(1):65-71
references.pmid: 21194951
references.citation: Biomaterials. 2011 Dec;32(34):9021-30
references.pmid: 21889204
references.citation: Hum Mol Genet. 2006 Jun 1;15(11):1894-913
references.pmid: 16644866
references.citation: Nat Methods. 2012 Jul;9(7):671-5
references.pmid: 22930834
references.citation: Dev Biol. 2004 Jun 15;270(2):488-98
references.pmid: 15183728
references.citation: Dev Cell. 2003 Aug;5(2):351-8
references.pmid: 12919684
references.citation: Nature. 2010 Sep 30;467(7315):596-9
references.pmid: 20823850
references.citation: Proc Natl Acad Sci U S A. 2011 May 10;108(19):7950-5
references.pmid: 21498687
references.citation: J Cell Biol. 2001 Apr 16;153(2):273-82
references.pmid: 11309409
references.citation: Lancet Oncol. 2012 Apr;13(4):e178-85
references.pmid: 22469128
references.citation: Nat Rev Genet. 2012 Apr 18;13(5):358-69
references.pmid: 22510765
references.citation: Nat Rev Cancer. 2009 Jan;9(1):28-39
references.pmid: 19104514
references.citation: Cancer Res. 2010 Nov 1;70(21):8736-47
references.pmid: 20959481
references.citation: N Engl J Med. 2011 Jun 30;364(26):2507-16
references.pmid: 21639808
references.citation: Transplant Proc. 2005 Jan-Feb;37(1):525-6
references.pmid: 15808698
references.citation: Cancer J. 2011 Nov-Dec;17(6):505-11
references.pmid: 22157295
references.citation: J Clin Endocrinol Metab. 2008 Jun;93(6):2194-201
references.pmid: 18381570
references.citation: Mol Cancer. 2006 Dec 02;5:67
references.pmid: 17140455
references.citation: Nature. 2001 Nov 1;414(6859):105-11
references.pmid: 11689955
references.citation: Nature. 2009 Oct 1;461(7264):649-3
references.pmid: 19718018
references.citation: J Cell Biol. 2000 Mar 6;148(5):849-56
references.pmid: 10704436
references.citation: Stem Cells Dev. 2007 Dec;16(6):1003-16
references.pmid: 18004940
references.citation: Cell. 2011 Aug 19;146(4):633-44
references.pmid: 21854987
references.citation: Biochem Biophys Res Commun. 2012 May 25;422(1):80-4
references.pmid: 22560902
references.citation: N Engl J Med. 2014 Nov 13;371(20):1867-76
references.pmid: 25265494
references.citation: Proc Natl Acad Sci U S A. 2007 Jun 19;104(25):10465-9
references.pmid: 17563371
references.citation: Adv Pathobiol. 1977;(6):141-52
references.pmid: 899956
references.citation: Cancer Cell. 2010 Dec 14;18(6):683-95
references.pmid: 21156289
references.citation: J Cell Physiol. 2010 Nov;225(2):417-28
references.pmid: 20506199
references.citation: Bioinformatics. 2004 Jun 12;20(9):1464-5
references.pmid: 14962934
references.citation: Clin Cancer Res. 2008 Jan 1;14(1):230-9
references.pmid: 18172275
references.citation: Int J Surg Pathol. 2005 Jan;13(1):1-8
references.pmid: 15735849
references.citation: J Transl Med. 2009 Mar 23;7:20
references.pmid: 19309508
references.citation: Oncogene. 2012 Nov 22;31(47):4898-911
references.pmid: 22286766
references.citation: Trends Pharmacol Sci. 2002 Jan;23 (1):40-5
references.pmid: 11804650
references.citation: J Biomed Inform. 2011 Oct;44(5):839-47
references.pmid: 21605702
references.citation: Nature. 2010 Dec 16;468(7326):973-7
references.pmid: 21107323
references.citation: Br J Cancer. 2009 Feb 10;100(3):431-5
references.pmid: 19156138
references.citation: Genes Dev. 2011 Oct 15;25(20):2173-86
references.pmid: 22012620
references.citation: Development. 2005 Nov;132(21):4653-62
references.pmid: 16224045