The study of iodide uptake change by transfection of a recombinant eukaryotic expression plasmid pcDNA3.1/hTSHR cDNA in FTC-133 cell lines

Objectives To investigate the changes of iodide uptake by transfection of a recombinant eukaryotic expression plasmid pcDNA3.1/hTSHR-cDNA in FTC-133 cell lines.

Methods The recombinant plasmid pcDNA3.1/hTSHR was verified by restriction enzyme digestion and DNA sequencing. Then it was transfected into FTC-133 cell lines by Lipofectin method. Target protein expression was detected by immunofluorescence and the mRNAs were detected by real-time PCR. Radioiodine uptake was measured with a gamma counter. Statistical analyses was performed by t-test analysis using SPSS 13.0 software. A value of P <0.05 was considered significant.

Results The sequence analysis confirmed that pcDNA3.1/hTSHR had been constructed successfully. After transfection with a recombinant plasmid pcDNA3.1/hTSHR cDNA, expression of the hTSHR protein in the FTC-133 was detected by immunofluorescence which was localized at the cell surface and cytoplasm. Radioiodine uptake of experiment group after infection was 2.9 times more (t=28.63, P<0.01) than that of control group. mRNAs of TSHR, NIS, TPO and Tg were elevated by 1.74 times (t=5.959, P<0.01),7.2 times(t=3.807, P<0.05), 2.88 times(t=4.769, P<0.01), 2.67 times (t= 6.388, P<0.01) respectively comparison with control group.

Conclusions Radioiodide uptake was triggered by transfection of the recombinant plasmid pcDNA3.1/hTSHR in FTC-133 cell lines. It outlined the potential of this novel gene therapy approach for radiotherapy of thyriod cancer.

Research Support This work was supported by Shanghai Leading Academic Discipline Project