title: Epithelial-mesenchymal transition markers in lymph node metastases and primary breast tumors - relation to dissemination and proliferation.
abstract: Epithelial-mesenchymal transition (EMT) was shown to enhance metastatic abilities of cancer cells, but it remains elusive in clinical samples. Moreover, EMT is rarely studied in lymph node metastases (LNM), thus limiting our understanding of its role outside of the primary tumors (PT). We collected a set of samples including triplets - PT, circulating tumor cells (CTCs)-enriched blood samples and LNM from 108 early breast cancer patients. With immunohistochemistry we analyzed levels of EMT effectors - E-cadherin, vimentin and N-cadherin in LNM, central areas and margins of PT. Additionally, expression of EMT core regulators TWIST1, SNAI1, SNAI2 was measured with RT-qPCR. Patients with E-cadherin loss had CTCs in 45% of the cases in comparison to 23% with normal E-cadherin level (P = 0.05). Mesenchymal phenotype of CTCs-enriched blood fractions was five-times more frequent in patients with E-cadherin loss in PT compared to PT with normal E-cadherin levels (P = 0.01). Epithelial/mesenchymal status of matched samples at different stages of dissemination was frequently discordant, especially for pairs involving CTCs, indicating high plasticity of tumor cells. LNM showed increased expression of TWIST1, SNAI1, SNAI2 accompanied by decreased Ki67 labeling index, with median Ki67 of 15% in PT and 10% in LNM (P = 0.0002). Our findings demonstrate that E-cadherin loss, not only in PT margin, might lead to seeding of especially malignant CTCs with mesenchymal phenotype. In comparison to PT, cells in LNM re-express E-cadherin, upregulate EMT transcription factors and reduce cell division rate, which could be viewed as their long-term survival strategy.
authors: Department of Medical Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk Dębinki 1 St, 80-211 Gdańsk, Poland ; Postgraduate School of Molecular Medicine, Medical University of Warsaw Żwirki i Wigury 61 St, 02-091 Warsaw, Poland.
authors: Aleksandra
authors: A
authors: Markiewicz
authors: Department of Oncology and Radiotherapy, Medical University of Gdańsk Dębinki 7 St, 80-211 Gdańsk, Poland.
authors: Marzena
authors: M
authors: Wełnicka-Jaśkiewicz
authors: Bank of Frozen Tissues and Genetic Specimens, Department of Medical Laboratory Diagnostics, Medical University of Gdańsk Dębinki 7 St, 80-211 Gdańsk, Poland.
authors: Barbara
authors: B
authors: Seroczyńska
authors: Bank of Frozen Tissues and Genetic Specimens, Department of Medical Laboratory Diagnostics, Medical University of Gdańsk Dębinki 7 St, 80-211 Gdańsk, Poland ; Department of Surgical Oncology, Medical University of Gdańsk M. Smoluchowskiego 17 St, 80-214 Gdańsk, Poland.
authors: Jarosław
authors: J
authors: Skokowski
authors: Department of Pathomorphology, Medical University of Gdańsk M. Smoluchowskiego 17 St, 80-214 Gdańsk, Poland.
authors: Hanna
authors: H
authors: Majewska
authors: Department of Pathomorphology, Medical University of Gdańsk M. Smoluchowskiego 17 St, 80-214 Gdańsk, Poland.
authors: Jolanta
authors: J
authors: Szade
authors: Department of Medical Biotechnology, Intercollegiate Faculty of Biotechnology, University of Gdańsk and Medical University of Gdańsk Dębinki 1 St, 80-211 Gdańsk, Poland ; Innovation Synergy Foundation Dragana 20/6, 80-807 Gdańsk, Poland.
authors: Anna J
authors: AJ
authors: Żaczek
keywords: Epithelial-mesenchymal transition; Ki67; circulating tumor cells; epithelial-mesenchymal plasticity; lymph node metastases
references: J Transl Med. 2012 Nov 19;10:226
references: 23157797
references: Oncogene. 2009 Nov 12;28(45):4022-33
references: 19718050
references: Cancer Microenviron. 2012 Apr;5(1):19-28
references: 21892699
references: Cell Cycle. 2009 Sep 15;8(18):2901-6
references: 19713745
references: Pathobiology. 2012;79(4):195-208
references: 22488297
references: Virchows Arch. 2011 Jul;459(1):31-9
references: 21638011
references: Genes Dev. 2013 Oct 15;27(20):2192-206
references: 24142872
references: Clin Cancer Res. 2012 Feb 15;18(4):993-1003
references: 22228641
references: Ann Oncol. 2014 Aug;25(8):1506-16
references: 24651410
references: Breast Cancer Res. 2003;5(6):R217-22
references: 14580257
references: Cancer Res. 2000 Jan 15;60(2):298-304
references: 10667580
references: Cell. 2009 Dec 24;139(7):1315-26
references: 20064377
references: Histopathology. 2005 Jun;46(6):685-93
references: 15910600
references: Cell. 2009 Nov 25;139(5):871-90
references: 19945376
references: Int J Cancer. 2012 Feb 15;130(4):808-16
references: 21387303
references: Am J Clin Pathol. 1995 Jul;104(1):42-9
references: 7611179
references: Am J Clin Pathol. 1998 Apr;109(4):431-8
references: 9535397
references: Cancer Cell. 2011 Nov 15;20(5):576-90
references: 22094253
references: Appl Immunohistochem Mol Morphol. 2012 Mar;20(2):103-7
references: 22553809
references: Nucleic Acids Res. 2013 Jul;41(Web Server issue):W562-8
references: 23761446
references: Breast Cancer Res Treat. 2002 Sep;75(1):1-3
references: 12500929
references: Cancers (Basel). 2013 Nov 08;5(4):1485-503
references: 24217115
references: Mol Biol Cell. 2007 May;18(5):1943-52
references: 17344479
references: Breast Cancer Res Treat. 2013 Feb;138(1):47-57
references: 23338761
references: Cancer Res. 2007 Dec 15;67(24):11721-31
references: 18089802
references: Ann Oncol. 2011 Aug;22(8):1736-47
references: 21709140
references: Exp Mol Pathol. 2013 Feb;94(1):168-73
references: 22985790
references: APMIS. 2007 Jan;115(1):52-65
references: 17223851
references: Ann Surg Oncol. 2010 Nov;17(11):3059-67
references: 20563657
references: Anticancer Res. 2007 Jul-Aug;27(4A):1969-74
references: 17649807
references: Hum Pathol. 2013 Jan;44(1):39-46
references: 22939959
references: J Mammary Gland Biol Neoplasia. 2012 Dec;17(3-4):241-9
references: 23011603
references: J Surg Oncol. 2013 Feb;107(2):188-94
references: 22886823
references: Cancer Epidemiol Biomarkers Prev. 2014 Jul;23(7):1164-75
references: 24793954
references: PLoS One. 2014 Apr 07;9(4):e93901
references: 24709997
references: Mol Cancer. 2010 Jul 07;9:179
references: 20609236
references: Breast Dis. 2008;29:27-36
references: 19029622
references: J Cancer Res Clin Oncol. 2015 Feb;141(2):189-201
references: 24965746
references: Virchows Arch. 2007 Jul;451(1):11-8
references: 17554555
references: Histopathology. 2013 Apr;62(5):695-701
references: 23347178
references: Nature. 2013 Jan 24;493(7433):487-8
references: 23344357
references: Cancer Res. 2008 Feb 15;68(4):989-97
references: 18281472
references: J Cell Biol. 1999 Nov 1;147(3):631-44
references: 10545506
references: J Clin Invest. 2009 Jun;119(6):1429-37
references: 19487819
references: Anticancer Res. 2007 Nov-Dec;27(6B):3903-8
references: 18225549
references: Cancer Lett. 2013 Nov 28;341(1):56-62
references: 23830804
references: Cancer Treat Rev. 2012 Oct;38(6):689-97
references: 22118888
references: Oncologist. 2012;17(10):1246-55
references: 22871798
references: Clinics (Sao Paulo). 2011;66(10 ):1765-71
references: 22012049
references: Cancer Res. 2009 Sep 15;69(18):7135-9
references: 19738043
references: Prostate. 2013 Jun;73(8):813-26
references: 23280481
references: Clin Exp Metastasis. 2011 Jan;28(1):55-63
references: 20963473
references: J Cell Sci. 2013 Jan 15;126(Pt 2):393-401
references: 23525005
references: Breast Cancer Res. 2009;11(4):R46
references: 19589136
references: Cancer Metastasis Rev. 2012 Dec;31(3-4):469-78
references: 22729277
references: Oncogene. 2012 Aug 16;31(33):3741-53
references: 22120722
references: J Pathol. 1997 Dec;183(4):404-11
references: 9496256
references: Genome Biol. 2007;8(2):R19
references: 17291332
references: Virchows Arch. 1997 Apr;430(4):285-9
references: 9134039
references: Genes Dev. 2004 May 15;18(10 ):1131-43
references: 15155580
references: Science. 2013 Feb 1;339(6119):580-4
references: 23372014
references: Stem Cell Reports. 2013 Dec 27;2(1):78-91
references: 24511467