Recovery of NIS expression in thyroid cancer cells by overexpression of Pax8 gene.
BACKGROUND
Recovery of iodide uptake in thyroid cancer cells by means of obtaining the functional expression of the sodium/iodide symporter (NIS) represents an innovative strategy for the treatment of poorly differentiated thyroid cancer. However, the NIS gene expression alone is not always sufficient to restore radioiodine concentration ability in these tumour cells.


METHODS
In this study, the anaplastic thyroid carcinoma ARO cells were stably transfected with a Pax8 gene expression vector. A quantitative RT-PCR was performed to assess the thyroid specific gene expression in selected clones. The presence of NIS protein was detected by Western blot and localized by immunofluorescence. A iodide uptake assay was also performed to verify the functional effect of NIS induction and differentiation switch.


RESULTS
The clones overexpressing Pax8 showed the re-activation of several thyroid specific genes including NIS, Pendrin, Thyroglobulin, TPO and TTF1. In ARO-Pax8 clones NIS protein was also localized both in cell cytoplasm and membrane. Thus, the ability to uptake the radioiodine was partially restored, associated to a high rate of efflux. In addition, ARO cells expressing Pax8 presented a lower rate of cell growth.


CONCLUSIONS
These finding demonstrate that induction of Pax8 expression may determine a re-differentiation of thyroid cancer cells, including a partial recovery of iodide uptake, fundamental requisite for a radioiodine-based therapeutic approach for thyroid tumours.
Presta
Ivan
I
Dipartimento di Medicina Sperimentale e Clinica G. Salvatore, University of Catanzaro Magna Graecia, Catanzaro, Italy. presta@unicz.it
Arturi
Franco
F
Ferretti
Elisabetta
E
Mattei
Tiziana
T
Scarpelli
Daniela
D
Tosi
Emanuele
E
Scipioni
Angela
A
Celano
Marilena
M
Gulino
Alberto
A
Filetti
Sebastiano
S
Russo
Diego
D
D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003593:Cytoplasm / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors; D006801:Humans; D007454:Iodides / Q000378:metabolism; D007457:Iodine Radioisotopes / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000096:biosynthesis; D008856:Microscopy, Fluorescence; D000072636:PAX8 Transcription Factor; D051761:Paired Box Transcription Factors / Q000096:biosynthesis*; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000076988:Sulfate Transporters; D027981:Symporters / Q000096:biosynthesis*; D013950:Thymus Gland / Q000378:metabolism; D013954:Thyroglobulin / Q000096:biosynthesis; D013964:Thyroid Neoplasms / Q000378:metabolism* / Q000532:radiotherapy; D014158:Transcription, Genetic; D014162:Transfection; D000099209:Sodium-Iodide Symporters
D017931:DNA Primers; D007454:Iodides; D007457:Iodine Radioisotopes; D026901:Membrane Transport Proteins; D000072636:PAX8 Transcription Factor; D051761:Paired Box Transcription Factors; D012313:RNA; D012333:RNA, Messenger; D011994:Recombinant Proteins; D000076988:Sulfate Transporters; D027981:Symporters; D013954:Thyroglobulin; D000099209:Sodium-Iodide Symporters; C083325:PAX8 protein, human; C085609:SLC26A4 protein, human