title: Glycosylation of Sodium/Iodide Symporter (NIS) Regulates Its Membrane Translocation and Radioiodine Uptake.
abstract: OBJECTIVE
  Human sodium/iodide symporter (hNIS) protein is a membrane glycoprotein that transports iodide ions into thyroid cells. The function of this membrane protein is closely regulated by post-translational glycosylation. In this study, we measured glycosylation-mediated changes in subcellular location of hNIS and its function of iodine uptake.
  
  
  METHODS
  HeLa cells were stably transfected with hNIS/tdTomato fusion gene in order to monitor the expression of hNIS. Cellular localization of hNIS was visualized by confocal microscopy of the red fluorescence of tdTomato. The expression of hNIS was evaluated by RT-PCR and immunoblot analysis. Functional activity of hNIS was estimated by radioiodine uptake. Cyclic AMP (cAMP) and tunicamycin were used to stimulate and inhibit glycosylation, respectively. In vivo images were obtained using a Maestro fluorescence imaging system.
  
  
  RESULTS
  cAMP-mediated Glycosylation of NIS resulted in increased expression of hNIS, stimulating membrane translocation, and enhanced radioiodine uptake. In contrast, inhibition of glycosylation by treatment with tunicamycin dramatically reduced membrane translocation of intracellular hNIS, resulting in reduced radioiodine uptake. In addition, our hNIS/tdTomato fusion reporter successfully visualized cAMP-induced hNIS expression in xenografted tumors from mouse model.
  
  
  CONCLUSIONS
  These findings clearly reveal that the membrane localization of hNIS and its function of iodine uptake are glycosylation-dependent, as our results highlight enhancement of NIS expression and glycosylation with subsequent membrane localization after cAMP treatment. Therefore, enhancing functional NIS by the increasing level of glycosylation may be suggested as a promising therapeutic strategy for cancer patients who show refractory response to conventional radioiodine treatment.
authors.affiliation: Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea.|Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.|Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Taemoon
authors.initials: T
authors.lastname: Chung
authors.affiliation: Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea.|Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.|Tumor Microenvironment Global Core Research Center, Seoul National University, Seoul, Korea.|Cancer Imaging Center, Seoul National University Hospital, Seoul, Korea.
authors.forename: Hyewon
authors.initials: H
authors.lastname: Youn
authors.affiliation: Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Chan Joo
authors.initials: CJ
authors.lastname: Yeom
authors.affiliation: Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea.|Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.|Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Keon Wook
authors.initials: KW
authors.lastname: Kang
authors.affiliation: Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul, Korea.|Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.|Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.|Tumor Microenvironment Global Core Research Center, Seoul National University, Seoul, Korea.
authors.forename: June-Key
authors.initials: JK
authors.lastname: Chung
mesh_terms: D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D007454:Iodides / Q000378:metabolism* / Q000627:therapeutic use; D007457:Iodine Radioisotopes / Q000627:therapeutic use*; D051379:Mice; D027981:Symporters / Q000235:genetics / Q000378:metabolism*; D013964:Thyroid Neoplasms / Q000235:genetics* / Q000473:pathology / Q000532:radiotherapy*; D014415:Tunicamycin / Q000008:administration & dosage; D023041:Xenograft Model Antitumor Assays; D000099209:Sodium-Iodide Symporters
chemical_list: D000242:Cyclic AMP; D007454:Iodides; D007457:Iodine Radioisotopes; D027981:Symporters; D014415:Tunicamycin; D000099209:Sodium-Iodide Symporters