title: Lectin RCA-I specifically binds to metastasis-associated cell surface glycans in triple-negative breast cancer.
abstract: BACKGROUND
  Triple-negative breast cancer (TNBC) patients often face a high risk of early relapse characterized by extensive metastasis. Previous works have shown that aberrant cell surface glycosylation is associated with cancer metastasis, suggesting that altered glycosylations might serve as diagnostic signatures of metastatic potential. To address this question, we took TNBC as an example and analyzed six TNBC cell lines, derived from a common progenitor, that differ in metastatic potential.
  
  
  METHODS
  We used a microarray with 91 lectins to screen for altered lectin bindings to the six TNBC cell lines. Candidate lectins were then verified by lectin-based flow cytometry and immunofluorescent staining assays using both TNBC/non-TNBC cancer cells. Patient-derived tissue microarrays were then employed to analyze whether the staining of Ricinus communis agglutinin I (RCA-I), correlated with TNBC severity. We also carried out real-time cell motility assays in the presence of RCA-I. Finally, liquid chromatography-mass spectrometry/tandem spectrometry (LC-MS/MS) was employed to identify the membrane glycoproteins recognized by RCA-I.
  
  
  RESULTS
  Using the lectin microarray, we found that the bindings of RCA-I to TNBC cells are proportional to their metastatic capacity. Tissue microarray experiments showed that the intensity of RCA-I staining is positively correlated with the TNM grades. The real-time cell motility assays clearly demonstrated RCA-I inhibition of adhesion, migration, and invasion of TNBC cells of high metastatic capacity. Additionally, a membrane glycoprotein, POTE ankyrin domain family member F (POTEF), with different galactosylation extents in high/low metastatic TNBC cells was identified by LC-MS/MS as a binder of RCA-I.
  
  
  CONCLUSIONS
  We discovered RCA-I, which bound to TNBC cells to a degree that is proportional to their metastatic capacities, and found that this binding inhibits the cell invasion, migration, and adhesion, and identified a membrane protein, POTEF, which may play a key role in mediating these effects. These results thus indicate that RCA-I-specific cell surface glycoproteins may play a critical role in TNBC metastasis and that the extent of RCA-I cell binding could be used in diagnosis to predict the likelihood of developing metastases in TNBC patients.
authors.forename: Shu-Min
authors.initials: SM
authors.lastname: Zhou
authors.forename: Li
authors.initials: L
authors.lastname: Cheng
authors.forename: Shu-Juan
authors.initials: SJ
authors.lastname: Guo
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Czajkowsky
authors.forename: Huafang
authors.initials: H
authors.lastname: Gao
authors.forename: Xiao-Fang
authors.initials: XF
authors.lastname: Hu
authors.forename: Sheng-Ce
authors.initials: SC
authors.lastname: Tao
mesh_terms: D002352:Carrier Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement; D005260:Female; D006801:Humans; D008562:Membrane Glycoproteins / Q000378:metabolism; D009362:Neoplasm Metastasis; D037121:Plant Lectins / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry; D064726:Triple Negative Breast Neoplasms / Q000378:metabolism* / Q000473:pathology*
chemical_list: D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D037121:Plant Lectins; D011134:Polysaccharides; C017707:Ricinus communis agglutinin-1
references.citation: Melanoma Res. 2004 Dec;14(6):479-85
references.pmid: 15577318
references.citation: Nat Rev Cancer. 2004 Feb;4(2):118-32
references.pmid: 14964308
references.citation: Nat Rev Cancer. 2005 Jul;5(7):526-42
references.pmid: 16069816
references.citation: Proc Natl Acad Sci U S A. 2006 Nov 21;103(47):17885-90
references.pmid: 17101985
references.citation: Nat Protoc. 2006;1(2):543-9
references.pmid: 17406280
references.citation: Invest Ophthalmol Vis Sci. 2008 Feb;49(2):743-50
references.pmid: 18235023
references.citation: Cancer Biol Ther. 2008 Jan;7(1):3-9
references.pmid: 18059174
references.citation: Glycobiology. 2008 Oct;18(10):761-9
references.pmid: 18625848
references.citation: Proc Natl Acad Sci U S A. 2009 Jun 9;106(23):9385-90
references.pmid: 19458257
references.citation: Genes Dev. 2009 Aug 15;23(16):1882-94
references.pmid: 19608765
references.citation: BMC Cancer. 2009;9:357
references.pmid: 19811634
references.citation: Nat Rev Clin Oncol. 2010 Feb;7(2):79-80
references.pmid: 20118978
references.citation: J Proteome Res. 2010 May 7;9(5):2565-72
references.pmid: 20235609
references.citation: Gan To Kagaku Ryoho. 2010 Jun;37(6):1113-6
references.pmid: 20567118
references.citation: Semin Cancer Biol. 2010 Jun;20(3):169-77
references.pmid: 20452433
references.citation: Glycoconj J. 2010 Aug;27(6):571-82
references.pmid: 20721622
references.citation: N Engl J Med. 2010 Nov 11;363(20):1938-48
references.pmid: 21067385
references.citation: Nat Rev Clin Oncol. 2010 Dec;7(12):683-92
references.pmid: 20877296
references.citation: Arch Surg. 2011 Jan;146(1):33-4
references.pmid: 21305708
references.citation: Mol Cell Proteomics. 2011 Jun;10(6):M110.007294
references.pmid: 21447706
references.citation: Breast Cancer Res. 2011;13(2):R35
references.pmid: 21457545
references.citation: Comb Chem High Throughput Screen. 2011 Sep;14(8):711-9
references.pmid: 21933110
references.citation: J Proteome Res. 2012 Feb 3;11(2):829-38
references.pmid: 22149100
references.citation: Mol Hum Reprod. 2012 Apr;18(4):204-15
references.pmid: 22053055
references.citation: Glycobiology. 2012 Jun;22(6):839-48
references.pmid: 22322011
references.citation: Oncogene. 2012 Sep 27;31(39):4317-32
references.pmid: 22179837
references.citation: PLoS One. 2012;7(10):e46536
references.pmid: 23094027
references.citation: Zhonghua Yi Xue Za Zhi. 2012 Nov 13;92(42):3001-3
references.pmid: 23328295
references.citation: J Pharm Pharmacol. 2013 Mar;65(3):411-22
references.pmid: 23356850
references.citation: Breast Cancer. 2013 Apr;20(2):191-3
references.pmid: 20082162
references.citation: Chem Soc Rev. 2013 May 21;42(10):4443-58
references.pmid: 23443201
references.citation: Cancer Cell. 2013 May 13;23(5):573-81
references.pmid: 23680145
references.citation: Mol Cancer. 2013;12:63
references.pmid: 23767917
references.citation: PLoS One. 2013;8(9):e72011
references.pmid: 24023725
references.citation: Breast. 2013 Oct;22(5):958-63
references.pmid: 23756383
references.citation: Proteomics Clin Appl. 2013 Oct;7(9-10):677-89
references.pmid: 23775902
references.citation: J Cancer Res Clin Oncol. 2001 Apr;127(4):231-6
references.pmid: 11315257
references.citation: Cancer Cell. 2003 Jun;3(6):537-49
references.pmid: 12842083
references.citation: Clin Chem Lab Med. 2005;43(4):361-9
references.pmid: 15899650