title: Role of tumor cell surface lysosome-associated membrane protein-1 (LAMP1) and its associated carbohydrates in lung metastasis.
abstract: OBJECTIVE
  Expression of lysosome-associated membrane protein-1 (LAMP1) on the surface correlates with metastatic potential of B16 melanoma cells. Downregulation of their expression in high metastatic (B16F10) cells reduced their surface expression and metastatic potential. Present investigations explore if overexpression of LAMP1 on the surface of low metastatic (B16F1) cells augment their metastatic ability, and if so, how?
  
  
  METHODS
  B16F1 cells were transduced with lentiviral vector carrying mutant-LAMP1 (Y386A) (mutLAMP1). Surface expression of LAMP1 and carbohydrates was analyzed by flow cytometry, immunofluorescence and/or immunoprecipitation and Western blotting. Cell spreading and motility were assessed on components of extracellular matrix (ECM) (fibronectin) and basement membrane (BM) (matrigel), and galectin-3-coated coverslips/plates. Metastatic potential was assessed using experimental metastasis assay.
  
  
  RESULTS
  Pre-incubation with anti-LAMP1 antibodies significantly reduced lung metastasis of B16F10 cells. Overexpression of mutLAMP1 significantly increased its surface expression on B16F1 cells, resulting in increased cellular spreading and motility on fibronectin and matrigel. LAMP1 is the major carrier of poly-N-acetyllactosamine (polyLacNAc) on B16F10 cells. However, significantly higher expression of mutLAMP1 had no effect on galectin-3 binding on cell surface or on spreading or motility of cells on galectin-3-coated coverslips/plates. These cells also failed to show any gain in metastatic ability. This could be because LAMP1 from these cells carried significantly lower levels of polyLacNAc in comparison with B16F10 cells.
  
  
  CONCLUSIONS
  PolyLacNAc on B16F10 cells and galectin-3 on lungs are the major participants in melanoma metastasis. Although surface LAMP1 promotes interactions with organ ECM and BM, carbohydrates on LAMP1 play a decisive role in dictating lung metastasis.
authors.affiliation: Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Tata Memorial Centre, Sector 22, Kharghar, Navi Mumbai, 410210, India.
authors.forename: Akhil Kumar
authors.initials: AK
authors.lastname: Agarwal
authors.forename: Nithya
authors.initials: N
authors.lastname: Srinivasan
authors.forename: Rashmi
authors.initials: R
authors.lastname: Godbole
authors.forename: Shyam K
authors.initials: SK
authors.lastname: More
authors.forename: Srikanth
authors.initials: S
authors.lastname: Budnar
authors.forename: Rajiv P
authors.initials: RP
authors.lastname: Gude
authors.forename: Rajiv D
authors.initials: RD
authors.lastname: Kalraiya
mesh_terms: D000818:Animals; D050260:Carbohydrate Metabolism; D002465:Cell Movement / Q000502:physiology; D005260:Female; D037502:Galectin 3 / Q000378:metabolism; D008175:Lung Neoplasms / Q000378:metabolism* / Q000556:secondary*; D051907:Lysosomal Membrane Proteins / Q000378:metabolism*; D008546:Melanoma, Experimental / Q000378:metabolism* / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D014407:Tumor Cells, Cultured; D052118:Lysosomal-Associated Membrane Protein 1
chemical_list: D037502:Galectin 3; D051907:Lysosomal Membrane Proteins; C052255:Lamp1 protein, mouse; D052118:Lysosomal-Associated Membrane Protein 1