title: Efficacy of lentiviral-mediated transfection of hTSHR in poorly differentiated thyroid carcinoma cell line.
abstract: BACKGROUND
  Dedifferentiated thyroid cancer is often incurable because it does not respond to radioiodine. This study aimed to investigate iodide uptake and the expressions of thyroid-specific molecules after the transfection of human thyrotropin receptor (hTSHR) gene in poorly differentiated follicular thyroid cancer cell line (FTC-133).
  
  
  METHODS
  pGC-FU-hTSHR-GFP-lentivirus and pGC-FU-GFP-lentivirus were added into FTC-133 cells respectively. The parental cells were defined as the blank group. Cells transduced with pGC-FU-GFP and pGC-FU-hTSHR-GFP were defined as the control group and experimental group respectively. The efficiency of transfection was observed under a fluorescence microscope. (125)I uptake by FTC-133 was analyzed by measuring the radioactivity. Real time-PCR, western blotting and radioimmunoassay were applied to detect the expressions of mRNAs and proteins of Na(+)/I(-) symporter (NIS), thyroid-stimulating hormone receptor (TSHR), thyroid peroxidase (TPO) and thyroglobulin (Tg) in FTC-133.
  
  
  RESULTS
  The green fluorescence was present in 80% of the transduced cells under fluorescence microscope. The iodine uptake of cells transduced with pGC-FU-TSHR-GFP was 3.3 times higher than that in the other two groups (P<0.01). NIS, TSHR, TPO and Tg had been significantly up-regulated in the experimental group as compared to the control group (P<0.01) and the blank group (P<0.01).
  
  
  CONCLUSIONS
  The hTSHR transfection in FTC-133 improved the expression of thyroid-specific molecules including TSHR, NIS, TPO and Tg and radioiodide uptake.
authors.affiliation: Department of Nuclear Medicine, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Cheng
authors.forename: Hongliang
authors.initials: H
authors.lastname: Fu
authors.forename: Fang
authors.initials: F
authors.lastname: Feng
authors.forename: Chao
authors.initials: C
authors.lastname: Ma
authors.forename: Jianing
authors.initials: J
authors.lastname: Li
authors.forename: Suyun
authors.initials: S
authors.lastname: Chen
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
mesh_terms: D001692:Biological Transport / Q000235:genetics; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics; D007457:Iodine Radioisotopes / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics*; D009928:Organ Specificity; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011989:Receptors, Thyrotropin / Q000235:genetics*; D027981:Symporters / Q000235:genetics; D013954:Thyroglobulin / Q000235:genetics; D013964:Thyroid Neoplasms / Q000235:genetics / Q000473:pathology*; D014162:Transfection / Q000379:methods*; D000099209:Sodium-Iodide Symporters
chemical_list: D007453:Iodide Peroxidase; D007457:Iodine Radioisotopes; D012333:RNA, Messenger; D011989:Receptors, Thyrotropin; D027981:Symporters; D013954:Thyroglobulin; D000099209:Sodium-Iodide Symporters