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Overexpression of α2,3sialyl T-antigen in breast cancer determined by miniaturized glycosyltransferase assays and confirmed using tissue microarray immunohistochemical analysis

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Abstract

Glycan structure alterations during cancer regulate disease progression and represent clinical biomarkers. The study determined the degree to which changes in glycosyltransferase activities during cancer can be related to aberrant cell-surface tumor associated carbohydrate structures (TACA). To this end, changes in sialyltransferase (sialylT), fucosyltransferase (fucT) and galactosyltransferase (galT) activity were measured in normal and tumor tissue using a miniaturized enzyme activity assay and synthetic glycoconjugates bearing terminal LacNAc Type-I (Galβ1-3GlcNAc), LacNAc Type-II (Galβ1-4GlcNAc), and mucin core-1/Type-III (Galβ1-3GalNAc) structures. These data were related to TACA using tissue microarrays containing 115 breast and 26 colon cancer specimen. The results show that primary human breast and colon tumors, but not adjacent normal tissue, express elevated β1,3GalT and α2,3SialylT activity that can form α2,3SialylatedType-IIIglycans (Siaα2-3Galβ1-3GalNAc). Prostate tumors did not exhibit such elevated enzymatic activities. α1,3/4FucT activity was higher in breast, but not in colon tissue. The enzymology based prediction of enhanced α2,3sialylated Type-III structures in breast tumors was verified using histochemical analysis of tissue sections and tissue microarrays. Here, the binding of two markers that recognize Galβ1-3GalNAc (peanut lectin and mAb A78-G/A7) was elevated in breast tumor, but not in normal control, only upon sialidase treatment. These antigens were also upregulated in colon tumors though to a lesser extent. α2,3sialylatedType-III expression correlated inversely with patient HER2 expression and breast metastatic potential. Overall, enzymology measurements of glycoT activity predict truncated O-glycan structures in tumors. High expression of the α2,3sialylated T-antigen O-glycans occur in breast tumors. A transformation from linear core-1 glycan to other epitopes may accompany metastasis.

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Abbreviations

GlycoT:

Glycosyltransferase

Galβ1-3GalNAc:

Type-III or core-1 type acceptors

Galβ1-4GlcNAc:

Type II or LacNAc type chain

GalT:

Galactosyltransferase

FT or FucT:

Fucosyltransferase

SialylT:

Sialyltransferase

ST3[Galβ1,3GalNAc]:

α2,3 sialyltransferase acting on Galβ1-3GalNAc

ST3/6[Galβ1,4GlcNAc]:

α2,3/6 sialyltransferase acting on Galβ1-4GlcNAc

Bn:

Benzyl

PAA:

Polyacrylamide

Φ-NO2 :

p-nitrophenol

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Acknowledgments

This work was supported by the National Institutes of Health [grant HL63014(SN), CA121294 (KLM) and HL103411(SN)] and Department of Defense [grant DODPC050420 (KLM)]].

Conflict of interest

We declare that we have no conflict of interest.

Author information

Authors and Affiliations

  1. Chemical and Biological Engineering, State University of New York, 906 Furnas Hall, Buffalo, NY, 14260, USA

    Shilpa A. Patil & Sriram Neelamegham

  2. Pathology, Roswell Park Cancer Institute, Buffalo, NY, 14263, USA

    Wiam Bshara & Carl Morrison

  3. Cancer Biology, Roswell Park Cancer Institute, Buffalo, NY, 14263, USA

    E. V. Chandrasekaran & Khushi L. Matta

  4. TumorEnd LLC, Baton Rouge, LA, 70803, USA

    Khushi L. Matta

  5. NY State Center for Excellence in Bioinformatics and Life Sciences, State University of New York, Buffalo, NY, 14260, USA

    Sriram Neelamegham

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  1. Shilpa A. Patil
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  2. Wiam Bshara
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Correspondence to Sriram Neelamegham.

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Patil, S.A., Bshara, W., Morrison, C. et al. Overexpression of α2,3sialyl T-antigen in breast cancer determined by miniaturized glycosyltransferase assays and confirmed using tissue microarray immunohistochemical analysis. Glycoconj J 31, 509–521 (2014). https://doi.org/10.1007/s10719-014-9548-4

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