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Poly N-acetyllactosamine substitutions on N- and not O-oligosaccharides or Thomsen–Friedenreich antigen facilitate lung specific metastasis of melanoma cells via galectin-3

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Abstract

Galectin-3 on vascular endothelium has been shown to facilitate lung specific metastasis. Metastatic variants of B16 melanoma were chosen to identify specific ligands that mediate lung colonization via galectin-3. Flow cytometry showed that, galectin-3 binding to cells correlates with surface expression of poly N-acetyllactosamine (polylacNAc) but not with other reported ligands, e.g. Thomsen-Friedenreich (T/Tn) antigen. Immobilized galectin-3 promoted adhesion of melanoma cells in a metastasis dependent manner. Moreover, adhesion and galectin-3 binding to cells were specifically inhibited with lactose. These properties together with lung metastasis were inhibited with N-glycosylation inhibitor Swainsonine (SW), whereas, O-glycosylation inhibitor Benzyl-α-N-acetylgalactosamine (BG) had no effect. BG treatment significantly increased expression of T/Tn antigen on low metastatic cells; however, had no effect on their metastatic potential. The studies very comprehensively demonstrate the importance of polylacNAc substitutions on N-oligosaccharides in galectin-3 mediated lung metastasis.

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Acknowledgements

We thank Dr. Hakon Leffler, Lund’s University, Sweden, for the expression vector for rhGalectin-3 and National Centre for Cell Science, Pune for the melanoma cell lines. We acknowledge the help extended by Ms. Archana Upadhya for Western blots, Miss. Radhika Kamdar for Immunohistochemistry, Ms. Rekha Santani and Shamal Vetale for Flow Cytometry, Mrs. Sadhana Kannan for statistical analysis, Mr. Chavan, Pawar and Lokhande for technical help, Mr. Sawant and Shyam Chavan for the help in preparing illustrations. We acknowledge the financial assistance received from Department of Science and Technology, Government of India and Senior Research Fellowship to Miss. Nithya Srinivasan from Council for Scientific and Industrial Research, Government of India.

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  1. Advanced Centre for Treatment Research and Education in Cancer (ACTREC), Tata Memorial Centre, Sector 22, Kharghar, Navi Mumbai, 410210, India

    Nithya Srinivasan, Sanjay M. Bane, Shashikant D. Ahire, Arvind D. Ingle & Rajiv D. Kalraiya

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  1. Nithya Srinivasan
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Srinivasan, N., Bane, S.M., Ahire, S.D. et al. Poly N-acetyllactosamine substitutions on N- and not O-oligosaccharides or Thomsen–Friedenreich antigen facilitate lung specific metastasis of melanoma cells via galectin-3. Glycoconj J 26, 445–456 (2009). https://doi.org/10.1007/s10719-008-9194-9

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