Crosstalk Between Abnormal TSHR Signaling Activation and PTEN/PI3K in the Dedifferentiation of Thyroid Cancer Cells

Iodide uptake and the metabolism of thyroid cells are regulated by thyrotropin (TSH)-TSH receptor (TSHR) signaling. Thus, it is necessary to elevate serum TSH levels by T4 withdraw or rTSH administration to facilitate radioiodide (131I) therapy for differentiated thyroid cancer (DTC). However, non-iodide-avid metastases of DTC which is dedifferentiated do not respond to stimulation by high levels of TSH, suggesting abnormal TSH-TSHR signal transduction in cancer cells. In addition, PI3K/AKT/mTOR signaling activation has been shown to be associated with the dedifferentiated phenotype of thyroid cancer, but the mechanism remains elusive. Therefore, in this study, we aimed to explore the role of abnormal TSH-TSHR signaling activation in regulating iodide uptake and cell mobility in thyroid cancer and its relationship with PI3K/AKT/mTOR signaling. We found that in thyroid cancer cells, TSH binds TSHR coupled to the Gα12/13 protein and then activates RhoA through interacting with leukemia associated RhoA guanine exchange factor (LARG). This results in a promigration tumorigenic phenotype independent of canonical TSHR-GαS signaling that regulates the expression of molecules involved in iodine uptake and metabolism. We observed that signaling pathways downstream of Gα12/13 signaling were increased, while that of Gαs signaling was decreased in thyroid cancer cells undergoing dedifferentiation compared to control cells following stimulation with different levels of TSH. PI3K/AKT/mTOR signaling activation enhanced Gα12/13 signaling through increasing LARG levels but also inhibited the expression of molecules downstream of Gαs signaling, including thyroid-specific molecules, and iodide uptake. In summary, our results demonstrate the noncanonical activation of TSH-TSHR signaling and its role in increasing the cell mobility and dedifferentiation of thyroid cancer through crosstalk with PI3K/AKT/mTOR signaling.

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Phosphoinositide-3-kinase inhibition induces sodium/iodide symporter expression in rat thyroid cells and human papillary thyroid cancer cells

Journal of Endocrinology ◽

10.1677/joe-08-0333 ◽

2008 ◽

Vol 199 (2) ◽

pp. 243-252 ◽

Cited By ~ 68

Author(s):

Takahiko Kogai ◽

Saima Sajid-Crockett ◽

Lynell S Newmarch ◽

Yan-Yun Liu ◽

Gregory A Brent

Keyword(s):

Thyroid Cancer ◽

Cancer Cells ◽

Papillary Thyroid Cancer ◽

Pi3k Pathway ◽

Papillary Thyroid ◽

Iodide Uptake ◽

Thyroid Cells ◽

Pathway Inhibition ◽

Rat Thyroid ◽

Thyroid Cancer Cells

TSH stimulation of sodium iodide symporter (NIS) expression in thyroid cancer promotes radioiodine uptake and is required to deliver an effective treatment dose. Activation of the insulin/phosphoinositide-3-kinase (PI3K) signaling pathway in TSH-stimulated thyroid cells reduces NIS expression at the transcriptional level. We, therefore, investigated the effects of PI3K pathway inhibition on iodide uptake and NIS expression in rat thyroid cell lines and human papillary thyroid cancer cells. A PI3K inhibitor, LY294002, significantly enhanced iodide uptake in two rat thyroid cell lines, FRTL-5 and PCCL3. The induction of Nis mRNA by LY294002 occurred 6 h after treatment, and was abolished by a translation inhibitor, cycloheximide. Expression of the transcription factor, Pax8, which stimulates NIS expression, was significantly increased in PCCL3 cells after LY294002 treatment. Removal of insulin abrogated the stimulatory effects of LY294002 on NIS mRNA and protein expression, but not on iodide uptake. These findings suggest that PI3K pathway inhibition results in post-translational stimulation of NIS. Inhibition of the PI3K pathway also significantly increased iodide uptake (∼3.5-fold) in BHP 2–7 papillary thyroid cancer cells (Ret/PTC1 positive), engineered to constitutively express NIS. Pharmacological inhibition of Akt, a factor stimulated by the PI3K pathway, increased exogenous NIS expression in BHP 2–7 as was seen with LY294002, but not increase the endogenous NIS expression in FRTL-5 cells. PI3K pathway inhibition increases functional NIS expression in rat thyroid cells and some papillary thyroid cancer cells by several mechanisms. PI3K inhibitors have the potential to increase radioiodide accumulation in some differentiated thyroid cancer.

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OR28-03 Drug Repurposing Identifies Inhibitors of the Proteostasis Network to Augment Radioiodine Uptake in Combinatorial Approaches Targeting Thyroid Cancer

Journal of the Endocrine Society ◽

10.1210/jendso/bvaa046.2135 ◽

2020 ◽

Vol 4 (Supplement_1) ◽

Author(s):

Martin L Read ◽

Katie Brookes ◽

Alice Fletcher ◽

Caitlin E M Thornton ◽

Mohammed Alshahrani ◽

...

Keyword(s):

Thyroid Cancer ◽

Cancer Cells ◽

High Throughput ◽

High Throughput Screening ◽

Drug Repurposing ◽

Iodide Uptake ◽

Thyroid Cells ◽

Radioiodine Uptake ◽

Repurposed Drugs ◽

Thyroid Cancer Cells

Abstract New combinatorial drug strategies are urgently needed to improve radioiodine (RAI) uptake and efficiently ablate thyroid cancer cells, thereby reducing the risk of recurrent disease. Drug repurposing offers the promise of identifying already approved compounds capable of inducing sodium iodide symporter (NIS) function to enhance iodide uptake. However, a lack of thyroid cell-based assays amenable to high-throughput screening has limited progress. We utilised the mutated yellow fluorescent protein (YFP) as a surrogate biosensor of intracellular iodide and screened the Prestwick Chemical Library (1200 drugs; 95% approved) for quenching of YFP fluorescence. This allowed us to identify putative candidate drugs which increased iodide uptake >2 SD above mean. Categorisation of these revealed a high proportion of drugs that modulate the proteostasis network (19/48; ~40%), including key processes in protein homeostasis such as endoplasmic reticulum-associated protein degradation (ERAD) and autophagy. Secondary screening validated the activity of proteostasis modulators in enhancing iodide uptake after ranking 73 leading compounds based on their pharmacologic (AUC, EMAX and EC50) and specificity of response (NIS+ve vs NIS-ve YFP-thyroid cells) at ten different drug doses (0.1 to 50 μM). Of importance, several repurposed drugs (e.g. ebastine, Prestwick N, Prestwick C and clotrimazole) in combination with the HDAC inhibitor vorinostat induced a robust enhancement in RAI uptake in thyroid cancer cells (TPC-1 and 8505C NIS+ve cells, up to 11-fold vs DMSO, P<0.001), which was significantly greater than using vorinostat alone (up to 3-fold, P<0.01). For clotrimazole, we designed 7 new chemical derivatives, 3 of which showed enhanced aqueous solubility and retained the ability to significantly enhance RAI uptake. TaqMan RT-PCR revealed that, in contrast to vorinostat, our repurposed drugs failed to alter NIS mRNA expression, highlighting post-transcriptional mechanisms. Critically, 11 repurposed drugs induced significant gains in RAI uptake in human primary thyroid cells (up to 4.1-fold; P<0.05), the most physiological setting for NIS function. In conclusion, we performed high-throughput screening and identified proteostasis modulators, as well as other repurposed drugs, that markedly enhance radioiodine uptake. Further clinical investigation of these drugs might offer new combinatorial approaches, especially with existing therapies, to improve the treatment of thyroid cancer.

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Paracrine Effect of Human Chorionic Gonadotropin Ectopically Produced from Papillary Thyroid Cancer Cells on Growth and Function of FRTL-5 Rat Thyroid Cells

Thyroid ◽

10.1089/thy.1997.7.779 ◽

1997 ◽

Vol 7 (5) ◽

pp. 779-782 ◽

Cited By ~ 1

Author(s):

NORIKO SAKAGUCHI ◽

MASAYOSHI YOSHIMURA ◽

JEROME M. HERSHMAN ◽

MITSUSHIGE NISHIKAWA ◽

MITSUO INADA

Keyword(s):

Thyroid Cancer ◽

Cancer Cells ◽

Papillary Thyroid Cancer ◽

Chorionic Gonadotropin ◽

Human Chorionic Gonadotropin ◽

Papillary Thyroid ◽

Thyroid Cells ◽

Rat Thyroid ◽

And Function ◽

Thyroid Cancer Cells

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Dual Oncogenic/Anti-Oncogenic Role of PATZ1 in FRTL5 Rat Thyroid Cells Transformed by the Ha-RasV12 Oncogene

Genes ◽

10.3390/genes10020127 ◽

2019 ◽

Vol 10 (2) ◽

pp. 127 ◽

Cited By ~ 1

Author(s):

Michela Vitiello ◽

Giuseppe Palma ◽

Mario Monaco ◽

Anna Bello ◽

Simona Camorani ◽

...

Keyword(s):

Stem Cell ◽

Thyroid Cancer ◽

Cancer Cells ◽

Tumor Growth Rate ◽

Stem Cell Markers ◽

Thyroid Cells ◽

Mesenchymal Transition ◽

Rat Thyroid ◽

The Impact ◽

Thyroid Cancer Cells

PATZ1 is a transcriptional factor downregulated in thyroid cancer whose re-expression in thyroid cancer cells leads to a partial reversion of the malignant phenotype, including the capacity to proliferate, migrate, and undergo epithelial-to-mesenchymal transition. We have recently shown that PATZ1 is specifically downregulated downstream of the Ras oncogenic signaling through miR-29b, and that restoration of PATZ1 in Ha-Ras transformed FRTL5 rat thyroid cells is able to inhibit their capacities to proliferate and migrate in vitro. Here, we analyzed the impact of PATZ1 expression on the in vivo tumorigenesis of these cells. Surprisingly, FRTL5-Ras-PATZ1 cells showed enhanced tumor initiation when engrafted in nude mice, even if their tumor growth rate was reduced compared to that of FRTL5-Ras control cells. To further investigate the cause of the enhanced tumor engraftment of FRTL5-Ras-PATZ1 cells, we analyzed the stem-like potential of these cells through their capacity to grow as thyrospheres. The results showed that restoration of PATZ1 expression in these cells increases stem cell markers’ expression and self-renewal ability of the thyrospheres while limiting their growth capacity. Therefore, we suggest that PATZ1 may play a role in enhancing the stem cell potential of thyroid cancer cells, but, at the same time, it impairs the proliferation of non-stem cells.

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Spheroid formation of human thyroid cancer cells under simulated microgravity: a possible role of CTGF and CAV1

Cell Communication and Signaling ◽

10.1186/1478-811x-12-32 ◽

2014 ◽

Vol 12 (1) ◽

pp. 32 ◽

Cited By ~ 45

Author(s):

Elisabeth Warnke ◽

Jessica Pietsch ◽

Markus Wehland ◽

Johann Bauer ◽

Manfred Infanger ◽

...

Keyword(s):

Thyroid Cancer ◽

Cancer Cells ◽

Simulated Microgravity ◽

Human Thyroid ◽

Spheroid Formation ◽

Thyroid Cancer Cells

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The role of human achaete-scute homolog-1 in medullary thyroid cancer cells

Surgery ◽

10.1016/s0039-6060(03)00418-5 ◽

2003 ◽

Vol 134 (6) ◽

pp. 866-871 ◽

Cited By ~ 45

Author(s):

Rebecca S Sippel ◽

Jennifer E Carpenter ◽

Muthusamy Kunnimalaiyaan ◽

Herbert Chen

Keyword(s):

Thyroid Cancer ◽

Cancer Cells ◽

Medullary Thyroid Cancer ◽

Medullary Thyroid ◽

Thyroid Cancer Cells

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miR-32-5p Inhibits the Proliferation, Migration and Invasion of Thyroid Cancer Cells by Regulating Twist1

10.21203/rs.3.rs-552882/v1 ◽

2021 ◽

Author(s):

Qing Liu ◽

Ouyang Li ◽

Chi Zhou ◽

Yu Wang ◽

Chunxue He ◽

...

Keyword(s):

Thyroid Cancer ◽

Cancer Cells ◽

Epithelial Mesenchymal Transition ◽

Transition Process ◽

Underlying Mechanism ◽

Luciferase Assay ◽

Migration And Invasion ◽

Mesenchymal Transition ◽

Thyroid Cancer Cells

Abstract Background: Thyroid cancer is the most prevalent malignancy and one of the leading causes of cancer-related deaths. Recent studies have revealed that microRNAs (miRNAs) play an important role in tumorigenesis in various cancer types by affecting the expression of its targets. However, the role of miR-32-5p in thyroid cancer remains limited. Methods: In this study, we attempt to explore the role of miR-32-5p in thyroid cancer and elucidate the underlying mechanism. Expression of miR-32-5p was determined by quantitative reverse transcription PCR. Functional assays were performed by CCK-8 assay, cell colony assay, cell apoptosis assay, cell migration and invasion assays, cell cycle assay and luciferase assay. Protein expression was analyzed by Western blot.Results: In the present study, the role of miR-32-5p in thyroid cancer was firstly explored. It is found that miR-32-5p was downregulated in thyroid cancer tissues and cells. Overexpression of miR-32-5p inhibited thyroid cancer cells proliferation, migration, invasion and epithelial‐mesenchymal transition process; while suppression of miR-32-5p exhibited an opposite effect on thyroid cancer cells. In addition, In addition, a luciferase assay showed Twist1 was identified as a direct target of miR-32-5p in thyroid cancer, and further study showed that restoration of Twist1 attenuated the biological effect of miR-32-5p on thyroid cancer cells. Conclusion: In conclusion, our results demonstrated miR-32-5p functions as a tumor suppressor by targeting Twist1 in thyroid cancer, providing a novel insight into thyroid cancer therapy.

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AUF1 Promotes Proliferation and Invasion of Thyroid Cancer via Downregulation of ZBTB2 and Subsequent TRIM58

Frontiers in Oncology ◽

10.3389/fonc.2021.681736 ◽

2021 ◽

Vol 11 ◽

Author(s):

Xin Du ◽

Jia-Mei Wang ◽

Da-Lin Zhang ◽

Tong Wu ◽

Xiao-Yan Zeng ◽

...

Keyword(s):

Thyroid Cancer ◽

Cancer Cells ◽

Binding Sites ◽

Critical Role ◽

Common Type ◽

Migration And Invasion ◽

Papillary Thyroid ◽

Thyroid Cancer Cells ◽

Proliferation And Invasion

The pathogenesis of papillary thyroid cancer (PTC), the most common type of thyroid cancer, is not yet fully understood. This limits the therapeutic options for approximately 7% of invasive PTC patients. The critical role of AUF1 in the progression of thyroid cancer was first reported in 2009, however, its molecular mechanism remained unclear. Our study used CRISPR/Cas 9 system to knockdown AUF1 in IHH4 and TPC1 cells. We noticed that the expression of TRIM58 and ZBTB2 were increased in the AUF1 knockdown IHH4 and TPC1 cells. When TRIM58 and ZBTB2 were inhibited by small hairpin RNAs (shRNAs) against TRIM58 (shTRIM58) and ZBTB2 (shZBTB2), respectively, the proliferation, migration, and invasion ability of the AUF1-knockdown IHH4 and TPC1 cells were increased. In addition, two ZBTB2 binding sites (-719~-709 and -677~-668) on TRIM58 promoter and two AUF1 binding sites (1250-1256 and 1258-1265) on ZBTB2 3’-UTR were identified. These results suggested that AUF1 affecting thyroid cancer cells via regulating the expression of ZBTB2 and TRIM58.

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