Long-term culture and functional characterization of follicular cells from adult normal human thyroids.

Curcio, Francesco; Ambesi-Impiombato, Francesco Saverio; Perrella, Giuseppina; Coon, Hayden G.

doi:10.1073/pnas.91.19.9004

Cited by 85 publications

(83 citation statements)

References 21 publications

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“…H1/10P culture growth medium, a basic culture proliferation medium with composition similar to that previously used for human thyroid cells, was used (33,34) with and without 50 g/ml uridine to allow growth of cells with impaired mitochondrial function.…”

Section: Methodsmentioning

confidence: 99%

Disruptive mitochondrial DNA mutations in complex I subunits are markers of oncocytic phenotype in thyroid tumors

Gasparre

Porcelli²,

Bonora

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

262

215

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Oncocytic tumors are a distinctive class of proliferative lesions composed of cells with a striking degree of mitochondrial hyperplasia that are particularly frequent in the thyroid gland. To understand whether specific mitochondrial DNA (mtDNA) mutations are associated with the accumulation of mitochondria, we sequenced the entire mtDNA in 50 oncocytic lesions (45 thyroid tumors of epithelial cell derivation and 5 mitochondrion-rich breast tumors) and 52 control cases (21 nononcocytic thyroid tumors, 15 breast carcinomas, and 16 gliomas) by using recently developed technology that allows specific and reliable amplification of the whole mtDNA with quick mutation scanning. Thirteen oncocytic lesions (26%) presented disruptive mutations (nonsense or frameshift), whereas only two samples (3.8%) presented such mutations in the nononcocytic control group. In one case with multiple thyroid nodules analyzed separately, a disruptive mutation was found in the only nodule with oncocytic features. In one of the five mitochondrion-rich breast tumors, a disruptive mutation was identified. All disruptive mutations were found in complex I subunit genes, and the association between these mutations and the oncocytic phenotype was statistically significant (P ‫؍‬ 0.001). To study the pathogenicity of these mitochondrial mutations, primary cultures from oncocytic tumors and corresponding normal tissues were established. Electron microscopy and biochemical and molecular analyses showed that primary cultures derived from tumors bearing disruptive mutations failed to maintain the mutations and the oncocytic phenotype. We conclude that disruptive mutations in complex I subunits are markers of thyroid oncocytic tumors.oncocytic tumors ͉ heteroplasmy ͉ homoplasmy ͉ damaging mutation ͉ microenvironment

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Section: Methodsmentioning

confidence: 99%

Disruptive mitochondrial DNA mutations in complex I subunits are markers of oncocytic phenotype in thyroid tumors

Gasparre

Porcelli²,

Bonora

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

262

215

View full text Add to dashboard Cite

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“…4322288); DNA profiles were compared manually with the American Type Culture Collection (ATCC) database and to the DNA profiles reported by Schweppe and colleagues (21). Human primary culture of thyroid cells (P5) were obtained from F. Curcio (University Udine, Italy) in 2003 and passaged in our laboratory, as described (22), for fewer than 2 months after resuscitation. They were tested in 2010 by proliferation rate and expression of thyroid differentiation markers (TG, TPO, and TSHR).…”

Section: Cell Linesmentioning

confidence: 99%

CD44 Proteolysis Increases CREB Phosphorylation and Sustains Proliferation of Thyroid Cancer Cells

et al. 2012

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CD44 is a marker of cancer stem-like cells and epithelial-mesenchymal transition that is overexpressed in many cancer types, including thyroid carcinoma. At extracellular and intramembranous domains, CD44 undergoes sequential metalloprotease-and g-secretase-mediated proteolytic cleavage, releasing the intracellular protein fragment CD44-ICD, which translocates to the nucleus and activates gene transcription. Here, we show that CD44-ICD binds to the transcription factor CREB, increasing S133 phosphorylation and CREB-mediated gene transcription. CD44-ICD enhanced CREB recruitment to the cyclin D1 promoter, promoting cyclin D1 transcription and cell proliferation. Thyroid carcinoma cells harboring activated RET/PTC, RAS, or BRAF oncogenes exhibited CD44 cleavage and CD44-ICD accumulation. Chemical blockade of RET/PTC, BRAF, metalloprotease, or g-secretase were each sufficient to blunt CD44 processing. Furthermore, thyroid cancer cell proliferation was obstructed by RNA interference-mediated knockdown of CD44 or inhibition of g-secretase and adoptive CD44-ICD overexpression rescued cell proliferation. Together, these findings reveal a CD44-CREB signaling pathway that is needed to sustain cancer cell proliferation, potentially offering new molecular targets for therapeutic intervention in thyroid carcinoma. Cancer Res; 72(6);

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“…Transient transfections were effectuated with FUGENE reagent, as suggested by the manufacturer (Roche Molecular Diagnostics, Basel, Switzerland). P5 thyroid normal primary culture has been previously described (Curcio et al, 1994). Human papillary thyroid cancer cell lines TPC-1, FB2, NIM, BHP10-3, BHP17-10, BHP14-9, BC-PAP, BHP2-7, BHP5-16, have been described previously (Cerutti et al, 1996;Ohta et al, 2001;Basolo et al, 2002), and were maintained in DMEM supplemented with 10% foetal bovine serum, 1% penicillin-streptomicin, and 1% glutamine.…”

Section: Cell Lines and Plasmidsmentioning

confidence: 99%

RAI(ShcC/N-Shc)-dependent recruitment of GAB1 to RET oncoproteins potentiates PI3-K signalling in thyroid tumors

et al. 2005

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RAI, also named ShcC/N-Shc, one of the members of the Shc proteins family, is a substrate of the RET receptor tyrosine kinase. Here, we show that RAI forms a protein complex with both RET/MEN2A and RET/PTC oncoproteins. By coimmunoprecipitation, we found that RAI associates with the Grb2-associated binder1 (GAB1) adapter. This association is constitutive, but, in the presence of RET oncoproteins, both RAI and GAB1 are tyrosine-phosphorylated, and the stoichiometry of this interaction remarkably increases. Consequently, the p85 regulatory subunit of phosphatidylinositol-3 kinase (PI-3K) is recruited to the complex, and its downstream effector Akt is activated. We show that human thyroid cancer cell lines derived from papillary or medullary thyroid carcinoma (PTC or MTC) carrying, respectively, RET/PTC and RET/MEN2A oncoproteins express RAI proteins. We also show that human PTC samples express higher levels of RAI, when compared to normal thyroid tissue. In thyroid cells expressing RET/PTC1, ectopic expression of RAI protects cells from apoptosis; on the other hand, the silencing of endogenous RAI by small inhibitory duplex RNAs in a PTC cell line that expresses endogenous RET/PTC1, increases the rate of spontaneous apoptosis. These data suggest that RAI is a critical substrate for RET oncoproteins in thyroid carcinomas.

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Long-term culture and functional characterization of follicular cells from adult normal human thyroids.

Abstract: We have obtained long-term cultures of differentiated proliferating follicular cells from normal adult human thyroid gands.

Cited by 85 publications

References 21 publications

Disruptive mitochondrial DNA mutations in complex I subunits are markers of oncocytic phenotype in thyroid tumors

Disruptive mitochondrial DNA mutations in complex I subunits are markers of oncocytic phenotype in thyroid tumors

CD44 Proteolysis Increases CREB Phosphorylation and Sustains Proliferation of Thyroid Cancer Cells

RAI(ShcC/N-Shc)-dependent recruitment of GAB1 to RET oncoproteins potentiates PI3-K signalling in thyroid tumors

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