Application of the Cre/loxP System to Enhance Thyroid-Targeted Expression of Sodium/Iodide Symporter

Lin, Xiaoqin; Fischer, Andrew H.; Ryu, Kwon‐Yul; Cho, Je‐Yoel; Sferra, Thomas J.; Kloos, Richard T.; Mazzaferri, Ernest L.; Jhiang, Sissy

doi:10.1210/jc.2003-031963

Cited by 10 publications

(7 citation statements)

References 27 publications

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“…Adenoviral delivery also permits thyroid-targeted gene expression in primary human thyrocytes using the thyroglobulin promoter and a Cre/loxP system. (20). Given that cell imaging allows I Ϫ concentration to be monitored only in YFP-H148Q/I152L-expressing cells, a high transduction efficiency is needed to ensure that fluorescent cells are representative of the entire population.…”

Section: Advantages and Applications Of New Methodsmentioning

confidence: 99%

Cell-based imaging of sodium iodide symporter activity with the yellow fluorescent protein variant YFP-H148Q/I152L

Rhoden¹,

Cianchetta²,

Stivani³

et al. 2007

American Journal of Physiology-Cell Physiology

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The sodium iodide symporter (NIS) mediates iodide (I(-)) transport in the thyroid gland and other tissues and is of increasing importance as a therapeutic target and nuclear imaging reporter. NIS activity in vitro is currently measured with radiotracers and electrophysiological techniques. We report on the development of a novel live cell imaging assay of NIS activity using the I(-)-sensitive and genetically encodable yellow fluorescent protein (YFP) variant YFP-H148Q/I152L. In FRTL-5 thyrocytes stably expressing YFP-H148Q/I152L, I(-) induced a rapid and reversible decrease in cellular fluorescence characterized by 1) high affinity for extracellular I(-) (35 muM), 2) inhibition by the NIS inhibitor perchlorate, 3) extracellular Na(+) dependence, and 4) TSH dependence, suggesting that fluorescence changes are due to I(-) influx via NIS. Individual cells within a population of FRTL-5 cells exhibited a 3.5-fold variation in the rate of NIS-mediated I(-) influx, illustrating the utility of YFP-H148Q/I152L to detect cell-to-cell difference in NIS activity. I(-) also caused a perchlorate-sensitive decrease in YFP-H148Q/I152L fluorescence in COS-7 cells expressing NIS but not in cells lacking NIS. These results demonstrate that YFP-H148Q/I152L is a sensitive biosensor of NIS-mediated I(-) uptake in thyroid cells and in nonthyroidal cells following gene transfer and suggest that fluorescence detection of cellular I(-) may be a useful tool by which to study the pathophysiology and pharmacology of NIS.

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Section: Advantages and Applications Of New Methodsmentioning

confidence: 99%

Cell-based imaging of sodium iodide symporter activity with the yellow fluorescent protein variant YFP-H148Q/I152L

Rhoden¹,

Cianchetta²,

Stivani³

et al. 2007

American Journal of Physiology-Cell Physiology

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“…Recently, the stable transfection of the NIS gene over cells of follicular carcinoma (FTC-133) without NIS has been achieved, restoring the skills of 131 I uptake [84][85][86][87][88]. Recently, the stable transfection of the NIS gene over cells of follicular carcinoma (FTC-133) without NIS has been achieved, restoring the skills of 131 I uptake [84][85][86][87][88].…”

Section: Nis Gene Therapymentioning

confidence: 99%

Diagnostic Usefulness of Tumor Markers in the Thyroid Cytological Samples Extracted by Fine-Needle Aspiration Biopsy

Sanchis

2010

EMIDDT

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Thyroid nodules are quite common and present approximately in 10% of the population. Fine-needle aspiration biopsy has become the mainstay of thyroid nodule evaluation and the overall accuracy is excellent; however, some aspirates demonstrating indeterminate cytology results do not permit definitive diagnosis of malignancy, and in addition, there are no clear guidelines for the management of these lesions because the incidence of malignancy in indeterminate aspirates varies in the different studies published. In order to find molecular markers in an attempt to predict malignancy based on cytology, at least 70 molecular or cellular and genetic markers have been studied in thyroid nodules. This review focuses on some potential markers such as thyroid peroxidase, thyroglobulin, telomerase, galectin-3, RET/PTC and protein p53; some of them, such as thyroid peroxidases, thyroglobulin and galectin-3, can be studied in a routine pathology laboratory and are promising, but do not yet fulfil criteria required for their use in clinical practice. The American guidelines and the European consensus for the management of thyroid nodules and differentiated thyroid cancer do not recommend their systematic use because the evidence that they have provided is insufficient. On the other hand, information obtained through cytological smears permits the study of complex metabolic or genetic pathways, providing researchers with a high throughput tool to elucidate changes in the global expression patterns seen in tumour cells. This ability to take tumour biology into account would allow the selection of different drugs, considering the predominant altered pathways observed in these samples. Finally, all these data may provide the molecular groundwork for permitting future preoperative discrimination of follicular adenomas from hyperplastic nodules, and may ultimately guide therapeutic strategies.

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“…Recombinant adenovirus carrying LacZ or dominant negative MEK1 (A217/A221) was acquired from Cell Biolabs. Inc. Recombinant adenovirus carrying FLhNIS was engineered in our lab (Lin et al, 2004). For experiments shown in Fig 4, U0126 was added 24 h post-transduction.…”

Section: Methodsmentioning

confidence: 99%

MEK inhibition leads to lysosome-mediated Na+/I− symporter protein degradation in human breast cancer cells

Zhang¹,

Beyer²,

Jhiang³

2013

Endocrine-Related Cancer

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The Na+/I- symporter (NIS) is a transmembrane glycoprotein that mediates active iodide uptake into thyroid follicular cells. NIS-mediated iodide uptake in thyroid cells is the basis for targeted radionuclide imaging and treatment of differentiated thyroid carcinomas and their metastases. Furthermore, NIS is expressed in many human breast tumors but not in normal non-lactating breast tissue, suggesting that NIS-mediated radionuclide uptake may also allow the imaging and targeted therapy of breast cancer. However, functional cell surface NIS expression is often low in breast cancer, making it important to uncover signaling pathways that modulate NIS expression at multiple levels, from gene transcription to post-translational processing and cell surface trafficking. In this study, we investigated NIS regulation in breast cancer by MEK (MAPK/ERK kinase) signaling, an important cell signaling pathway involved in oncogenic transformation. We found that MEK inhibition decreased NIS protein levels in all-trans retinoic acid (tRA)/hydrocortisone treated MCF-7 cells as well as human breast cancer cells expressing exogenous NIS. The decrease in NIS protein levels by MEK inhibition was not accompanied by a decrease in NIS mRNA or a decrease in NIS mRNA export from the nucleus to the cytoplasm. NIS protein degradation upon MEK inhibition was prevented by lysosome inhibitors, but not by proteasome inhibitors. Interestingly, NIS protein level was correlated with MEK/ERK activation in human breast tumors from a tissue microarray. Taken together, MEK activation appears to play an important role in maintaining NIS protein stability in human breast cancers.

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Application of the Cre/loxP System to Enhance Thyroid-Targeted Expression of Sodium/Iodide Symporter

Cited by 10 publications

References 27 publications

Cell-based imaging of sodium iodide symporter activity with the yellow fluorescent protein variant YFP-H148Q/I152L

Cell-based imaging of sodium iodide symporter activity with the yellow fluorescent protein variant YFP-H148Q/I152L

Diagnostic Usefulness of Tumor Markers in the Thyroid Cytological Samples Extracted by Fine-Needle Aspiration Biopsy

MEK inhibition leads to lysosome-mediated Na+/I− symporter protein degradation in human breast cancer cells

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