Follicular Thyroid Carcinoma Cell Research Articles

Synchronous Primary Head and Neck Tumors: Follicular Thyroid Carcinoma and Squamous Cell Carcinoma of The Tonsil

Simultaneously occurring malignancies may be detected in different organs or tissues at any given time. Patients diagnosed with a tumor may be found to have another tumor or second primary cancer. Second primary cancers (SPCs) may be further classified as synchronous or metachronous. Synchronous SPCs are lesions detected simultaneously or within 6 months after the diagnosis of the primary tumor while metachronous SPCs are tumors diagnosed 6 months after primary tumor diagnosis.1 There is an increased risk of having second primary cancer in Head and Neck Squamous Cell Carcinoma (HNSCC) patients.1 In a study by Strojan et al. in 2013, among 2,106 head and neck cancer patients, 2.4% developed synchronous second primary cancers.2 A systematic review by Coco-Pelaz et al. in 2020, showed that second primary tumors most frequently occur in the head and neck area followed by the lungs and esophagus.3 We present a case of follicular thyroid carcinoma with an incidental finding of cervical lymph node metastatic squamous cell carcinoma from the tonsil and discuss the clinical presentation, ancillary procedures and management.

DICER1 syndrome and its various paediatric presentations: Case series and review of the literature

DICER1 syndrome is a rare tumour predisposition syndrome, associated with a range of benign and malignant tumours, which may occur during childhood. A high index of suspicion is required to ensure appropriate diagnosis and testing, with early treatment and surveillance of at-risk individuals. In this report, we present 5 patients with variants in DICER1 identified following diagnosis of a minimally invasive thyroid follicular cell carcinoma, a pineoblastoma, a pleuropulmonary blastoma, a urethral rhabdomyosarcoma and on sibling testing. Each of these children have presented at a young age, and 2 have presented with characteristic tumours prior to the ages currently recommended for initiation of routine screening. We discuss their presentation, management and follow up, as well as a review of the current literature on each associated tumour in relation to our patients. Overall, we demonstrate that DICER1 is a heterogenous condition and that there is a need for cascade testing of family members as well as regular screening for tumour development in affected children, although consideration should be made regarding initiating this screening at an earlier age depending on clinical findings.

On-the-fly Raman microscopy guaranteeing the accuracy of discrimination

Accelerating the measurement for discrimination of samples, such as classification of cell phenotype, is crucial when faced with significant time and cost constraints. Spontaneous Raman microscopy offers label-free, rich chemical information but suffers from long acquisition time due to extremely small scattering cross-sections. One possible approach to accelerate the measurement is by measuring necessary parts with a suitable number of illumination points. However, how to design these points during measurement remains a challenge. To address this, we developed an imaging technique based on a reinforcement learning in machine learning (ML). This ML approach adaptively feeds back "optimal" illumination pattern during the measurement to detect the existence of specific characteristics of interest, allowing faster measurements while guaranteeing discrimination accuracy. Using a set of Raman images of human follicular thyroid and follicular thyroid carcinoma cells, we showed that our technique requires 3,333 to 31,683 times smaller number of illuminations for discriminating the phenotypes than raster scanning. To quantitatively evaluate the number of illuminations depending on the requisite discrimination accuracy, we prepared a set of polymer bead mixture samples to model anomalous and normal tissues. We then applied a home-built programmable-illumination microscope equipped with our algorithm, and confirmed that the system can discriminate the sample conditions with 104 to 4,350 times smaller number of illuminations compared to standard point illumination Raman microscopy. The proposed algorithm can be applied to other types of microscopy that can control measurement condition on the fly, offering an approach for the acceleration of accurate measurements in various applications including medical diagnosis.

Spontaneous bilateral thyroid follicular cell carcinoma (subtype: compact cellular carcinoma) with C-cell complexes in a male beagle

Spontaneous bilateral thyroid follicular cell carcinoma (subtype: compact cellular carcinoma) with C-cell complexes in a male beagle

In Vitro Radioenhancement Using Ultrasound-Stimulated Microbubbles: A Comparison of Suspension and Adherent Cell States

Background: Ultrasound-stimulated microbubbles (USMB) have shown potential for enhancing radiation treatment via cavitation and sonoporation mechanisms. However, in vitro studies have produced inconsistent results, with adherent cells demonstrating no radioenhancement. This study aims to investigate the effect of cell adherence on in vitro radioenhancement using USMB and radiation. Method: Lung metastases of follicular thyroid carcinoma cells (FTC-238) and non-small cell lung carcinoma cells (NCI-H727) were treated, both when adhered and in suspension, using 1.6% (v/v) Definity™ microbubbles, ~90 s of 2 MHz ultrasound with mechanical index 0.9, and either 3 Gy or 6 Gy of megavoltage (MV) X-rays. The cell viability was measured using an MTS assay 72 h post-treatment, and statistical analysis was conducted using a three-way analysis of variance. Results: Statistically significant differences were observed for cells treated when adherent compared to suspended. An additive effect was detected in NCI-H727 cells treated in suspension, but not while adherent, while no enhancement was observed for FTC-238 cells in either culture state. Conclusions: To the best of our knowledge, this is the first study to directly compare the effect of cell adherence on the radioenhancement potential of USMB in vitro, and the first to do so using a metastatic cell line.

Piperlongumine Induces Cellular Apoptosis and Autophagy via the ROS/Akt Signaling Pathway in Human Follicular Thyroid Cancer Cells.

Thyroid cancer (TC) is the most common endocrine malignancy. Recently, the global incidence of TC has increased rapidly. Differentiated thyroid cancer includes papillary thyroid carcinoma (PTC) and follicular thyroid carcinoma (FTC), which are the most common types of TC. Although PTCs and FTCs exert good prognoses and high survival rates, FTCs tend to be more aggressive than PTCs. There is an urgent need to improve patient outcomes by developing effective therapeutic agents for FTCs. Piperlongumine exerts anti-cancer effects in various human carcinomas, including human anaplastic TCs and PTCs. However, the anti-cancer effects of piperlongumine in FTCs and the underlying mechanisms are yet to be elucidated. Therefore, in the present study, we evaluated the effect of piperlongumine on cell proliferation, cell cycle, apoptosis, and autophagy in FTC cells with flowcytometry and Western blot. We observed that piperlongumine caused growth inhibition, cell cycle arrest, apoptosis induction, and autophagy elevation in FTC cells. Activities of reactive oxygen species and the downstream PI3K/Akt pathway were the underlying mechanisms involved in piperlongumine mediated anti-FTC effects. Advancements in our understanding of the effects of piperlongumine in FTC hold promise for the development of novel therapeutic strategies.

Lipid droplet accumulation and adipophilin expression in follicular thyroid carcinoma

Follicular neoplasms of the thyroid include follicular thyroid carcinoma (FTC) and follicular thyroid adenoma (FTA). However, the differences in cytological findings between FTC and FTA remain undetermined. Here, we aimed to evaluate the accumulation of lipid droplets (LDs) and the expression of adipophilin (perilipin 2/ADRP/ADFP), a known LD marker, in cultured FTC cells. We also immunohistochemically compared adipophilin expression in the FTC and FTA of resected human thyroid tissues. Cultured FTC (FTC-133 and RO82W-1) possessed increased populations of LDs compared to thyroid follicular epithelial (Nthy-ori 3-1) cells. In vitro treatment with phosphatidylinositol-3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling inhibitors (LY294002, MK2206, and rapamycin) in FTC-133 cells downregulated the PI3K/Akt/mTOR/sterol regulatory element-binding protein 1 (SREBP1) signaling pathway, resulting in a significant reduction in LD accumulation. SREBP1 is a master transcription factor that controls lipid metabolism. Fluorescence immunocytochemistry revealed adipophilin expression in the LDs of FTC-133 cells. Immunohistochemical analysis of surgically resected human thyroid tissues revealed significantly increased expression of adipophilin in FTC compared with FTA and adjacent non-tumorous thyroid epithelia. Taken together, LDs and adipophilin were abundant in cultured FTC; the evaluation of adipophilin expression can help distinguish FTC from FTA in surgical specimens.

Ultrasound-Stimulated Microbubbles Enhance Radiation-Induced Cell Killing.

Recent in vivo studies using ultrasound-stimulated microbubbles as a localized radiosensitizer have had impressive results. While in vitro studies have also obtained similar results using human umbilical vein endothelial cells (HUVEC), studies using other cell lines have had varying results. This study was aimed at investigating any increases in radiation-induced cell killing in vitro using two carcinoma lines not previously investigated before (metastatic follicular thyroid carcinoma cells [FTC-238] and non-small cell lung carcinoma cells [NCI-H727]), in addition to HUVEC. Cells were treated using a combination of 1.6% (v/v) microbubbles, ∼90 s of 2-MHz ultrasound (mechanical index=0.8) and 0-6 Gy of kilovolt or MV X-rays. Cell viability assays obtained 72 h post-treatment were normalized to untreated controls, and analysis of variance was used to determine statistical significance. All cells treated with combined ultrasound-stimulated microbubbles and radiation exhibited decreased normalized survival, with statistically significant effects observed for the NCI-H727 cells. No statistically significant differences in effects were observed using kV compared with MV radiation. Further studies using increased microbubble concentrations may be required to achieve statistically significant results for the FTC-238 and HUVEC lines.

Silver nanoparticles induced apoptosis in papillary and follicular thyroid carcinoma cells

Uniform and Monodisperse AgNPs were synthesized via a simple chemical reduction approach the synthesized AgNPs' characterizations were as follows: UV–Vis analysis displayed Surface plasmon resonance (SPR) of AgNPs peaked around 425 nm. AgNPs were shown to have a crystalline structure with a face-centered cubic (FCC) lattice using X-ray diffraction (XRD) investigation. Transmission electron microscopy (TEM) images illustrated a spherical in shape and well dispersant AgNPs, having a 12 nm average size. The AgNPs' stability was confirmed by dynamic light scattering (DLS) data with the coincidence of hydrodynamic diameter to the nominal particle size obtained from TEM. Interestingly, the synthesized AgNPs demonstrated a dose-dependent reduction of thyroid carcinoma growth, the Papillary (MDA-T22) and the Follicular (FTC-133) cells with determined IC50 values of 71.35 and 52.79 μg/ml, respectively. Meanwhile, against normal cells (WRL-68) the AgNPs had no cytotoxic effect at low concentrations and minimal cytotoxicity at higher of 200–400 μg/ml with a significantly increased IC50 value reached to 218.4 μg ml−1. The induced apoptosis of AgNPs treated cancer cells was determined by a High-content screening (HCS) assay. Moreover, the apoptotic morphological changes were explored and the mechanism of affected living cells to apoptosis showed an increase in cell membrane permeability, cytochrome c level, and nuclear intensity in dose-dependent with a significant deferent at higher concentrations (100 and 200 μg mL−1) (p < 0.0001). Conversely, a noticeable decrease in cell viable count and mitochondrial membrane permeability was observed with AgNPs treatment compared with control. Our findings indicate the potential suitability of these nanoparticles for biological and clinical applications.

A recurrent somatic missense mutation in GNAS gene identified in familial thyroid follicular cell carcinomas in German longhaired pointer dogs

BackgroundWe previously reported a familial thyroid follicular cell carcinoma (FCC) in a large number of Dutch German longhaired pointers and identified two deleterious germline mutations in the TPO gene associated with disease predisposition. However, the somatic mutation profile of the FCC in dogs has not been investigated at a genome-wide scale.ResultsHerein, we comprehensively investigated the somatic mutations that potentially contribute to the inherited tumor formation and progression using high depth whole-genome sequencing. A GNAS p.A204D missense mutation was identified in 4 out of 7 FCC tumors by whole-genome sequencing and in 20 out of 32 dogs’ tumors by targeted sequencing. In contrast to this, in the human TC, mutations in GNAS gene have lower prevalence. Meanwhile, the homologous somatic mutation in humans has not been reported. These findings suggest a difference in the somatic mutation landscape between TC in these dogs and human TC. Moreover, tumors with the GNAS p.A204D mutation had a significantly lower somatic mutation burden in these dogs. Somatic structural variant and copy number alterations were also investigated, but no potential driver event was identified.ConclusionThis study provides novel insight in the molecular mechanism of thyroid carcinoma development in dogs. German longhaired pointers carrying GNAS mutations in the tumor may be used as a disease model for the development and testing of novel therapies to kill the tumor with somatic mutations in the GNAS gene.

Mixed medullary and follicular cell thyroid carcinoma in a dog.

A 12-y-old, castrated male Weimaraner dog was presented for a wellness examination. A 7-cm, firm mass was palpated on the left, ventral, mid-lateral neck. The neck mass was removed surgically and submitted for histopathology. A thyroid carcinoma was diagnosed based on microscopic examination. Immunohistochemistry for chromogranin-A, calcitonin, and thyroglobulin identified dual immunoreactivity of the latter two, and a final diagnosis was of a well-differentiated, compact, mixed medullary and follicular cell thyroid carcinoma. These neoplasms are rare in humans and have not been reported in dogs, to our knowledge.

Nav1.6 promotes the progression of human follicular thyroid carcinoma cells via JAK-STAT signaling pathway

Follicular thyroid carcinoma (FTC) is one of the most common malignant tumors of the endocrine system. Recent studies have shown that voltage-gated sodium channels (VGSCs) affect the proliferation, migration, and invasion of tumor cells. However, the expression and functions of VGSCs, and the molecular pathways activated by VGSCs in FTC cells remain unclear. Our studies revealed that the expression of Nav1.6, encoded by SCN8A, was the predominantly upregulated subtype of VGSCs in FTC tissues. Knockdown of Nav1.6 significantly inhibited the proliferation, epithelial–mesenchymal transition and invasiveness of FTC cells. Using gene set enrichment analysis and Kyoto Encyclopedia of Genes and Genomics, SCN8A was predicted to be related to the JAK-STAT signaling pathway. Hence, we targeted the JAK-STAT pathway and demonstrated that Nav1.6 enhanced FTC cell proliferation, epithelial–mesenchymal transition, and invasion by phosphorylating JAK2 to activate STAT3. Furthermore, downregulating the expression of Nav1.6 improve the susceptibility of FTC cells to ubenimex in vitro. These results suggest Nav1.6 accelerates FTC progression through JAK/STAT signaling and may be a potential target for FTC therapy.

Syntheses, characterization, and suppression efficiency of silver & silver iodide nanoparticle for proliferation, migration, and invasion in follicular thyroid carcinoma cells

In this study, a chemical co-precipitation method has been employed, silver iodide (AgI NPs) and silver nanoparticles (AgNPs) have been synthesized. UV–vis, FTIR, x-ray diffraction, FESEM, TEM, and other techniques have been used to examine the optical and structural properties of AgNPs and AgI NPs. The UV–vis absorption spectra gave the highest peak at 400 nm for AgNPs and AgI NPs at 434 nm. The x-ray data showed that the prepared AgNPs and AgI NPs were nanocrystalline cubic structures with crystallite sizes of 18 nm and 51 nm, respectively. The FESEM results show that synthesized AgNPs and AgI NPs agglomerate and aggregate. TEM data revealed that AgNPs have a quasi-spherical shape and Gaussian size distribution type. TEM analysis of AgI NPs with different magnifications revealed primarily spherical and well dispersed AgI NPs. TEM histogram shows that the particles were highly monodispersed AgNPs and AgI NPs with an average diameter of 11.5, 24.28 nm, respectively. According to the MTT assay results of FTC133cells, the cytotoxic action IC50 of AgNPs was (52.74 μg ml−1) and for AgI nanoparticles was (95.22 μg ml−1). It has been found that FTC133 cellular uptake was concentration, size- and time-dependent for both AgNPs and AgI NPs. The migrated FTC133 cell rates were reduced following AgNPs treatment to 75.7% and for AgI NPs treatment to 60% compared with the control group. Furthermore, Invasive FTC133 cell rates were reduced by 60% in the AgNPs treatment group and by 55.71 percent in the AgI NPs treatment group compared to the control group.

Follicular and Hurthle Cell Carcinoma: Comparison of Clinicopathological Features and Clinical Outcomes.

Background: Follicular thyroid carcinoma (FTC) and Hurthle cell carcinoma (HCC) are rare and aggressive thyroid cancers with limited published data comparing their outcomes or regarding their subtypes. The aim of this study was to describe clinicopathological features and compare clinical outcomes of patients with FTC and HCC based on the 2017 World Health Organization definition and extent of vascular invasion (VI). Methods: We retrospectively studied 190 patients with HCC and FTC primarily treated with surgery at Memorial Sloan Kettering Cancer Center between 1986 and 2015. Patients were classified as minimally invasive (MI), encapsulated angioinvasive with focal VI (EA-FVI), encapsulated angioinvasive with extensive VI (EA-EVI), and as widely invasive (WI). To compare clinical outcomes, patients were grouped as follows: group 1 = FTC-MI and FTC EA-FVI, group 2 = FTC EA-EVI and FTC-WI, group 3 = HCC-MI and HCC EA-FVI, group 4 = HCC EA-EVI and HCC-WI. Outcomes of interest were overall survival (OS), disease-specific survival (DSS), recurrence-free survival (RFS), locoregional recurrence-free survival (LRRFS), and distant recurrence-free survival (DRFS). Outcomes were determined using the Kaplan-Meier method and compared with log-rank test. Results: Patients with HCC (n = 111) were more likely to be older than 55 years old (59% vs. 27%, p < 0.001) with a tendency to present with more extensive VI (33% vs. 19%, p = 0.07) compared with FTC (n = 79). Comparing groups 1, 2, 3, and 4, group 4 patients were more likely to recur (DFS 98%, 93%, 98% vs. 73%, respectively, p = 0.0069). There was no statistically significant difference in OS, DSS LRRFS, or DRFS. Stratified by extent of VI (no, focal, and extensive VI), patients with extensive VI were more likely to recur (RFS 100%, 95%, 77%, p = 0.0025) and had poorer distant control (DRFS: 100%, 95%, 80%, p = 0.022), compared with patients absent or focal VI. Conclusions: Accurate assessment of the extent of VI and tumor phenotype (follicular vs. Hurthle) are essential in identifying patients at higher risk of recurrence.

Sophoridine Antiproliferative Effects in Human Thyroid Carcinoma Cells Mediated by Regulating PI3K/AKT/m-TOR Signalling Pathway

Present report determines the protective effect of Sophoridine against thyroid cancer. Follicular thyroid carcinoma cell line (WRO) and papillary thyroid carcinoma cell line (IHH-4) cancerous cells were treated with Sophoridine 0-200 µM and determine the cellular viability. Assessment of clonogenic assay, production of reactive oxygen species (ROS) was estimated and Sophoridine effect was also observed on apoptosis and PI3K/AKT/m-TOR signalling pathway using western blot assay. Sophoridine reduces the cellular viability, production of ROS and number of cell colonies in Sophoridine (50-200µM) treated group than Sophoridine 0µM. Treatment with Sophoridine ameliorates the apoptosis and PI3K/AKT/m-TOR pathway in WRO and IHH-4 cancerous cells. In conclusion, present investigation suggests the treatment with Sophoridine reduces the oxidative stress and enhances the apoptosis against thyroid cancer by regulating PI3K/AKT/m-TOR pathway.

Familial follicular cell thyroid carcinomas in a large number of Dutch German longhaired pointers.

Thyroid carcinomas (TCs) originating from follicular cells of the thyroid gland occur in both humans and dogs, and they have highly similar histomorphologic patterns. In dogs, TCs have not been extensively investigated, especially concerning the familial origin of TCs. Here, we report familial thyroid follicular cell carcinomas (FCCs) confirmed by histology in 54 Dutch origin German longhaired pointers. From the pedigree, 45 of 54 histopathologically confirmed cases are closely related to a pair of first‐half cousins in the past, indicating a familial disease. In addition, genetics contributed more to the thyroid FCC than other factors by an estimated heritability of 0.62 based on pedigree. The age of diagnosis ranged between 4.5 and 13.5 years, and 76% of cases were diagnosed before 10 years of age, implying an early onset of disease. We observed a significant higher pedigree‐based inbreeding coefficient in the affected dogs (mean F, 0.23) compared to unaffected dogs (mean F, 0.14), suggesting the contribution of inbreeding to tumour development. The unique occurrence of familial thyroid FCC in this dog population and the large number of affected dogs make this population an important model to identify the genetic basis of familial thyroid FCC in this breed and may contribute to the research into pathogenesis, prevention and treatment in humans.

ENDOG Impacts on Tumor Cell Proliferation and Tumor Prognosis in the Context of PI3K/PTEN Pathway Status.

Simple SummaryThe PI3K/AKT pathway is involved in cell survival and proliferation. Molecular aberrations and/or hyperactivation of the PI3K-PTEN-AKT axis are frequent in distinct cancer types such as endometrial carcinoma, the most common type of cancer of the female genital tract, and chronic lymphocytic leukemia (CLL), a mature B-cell neoplasm depending on B-cell receptor (BCR) activity, which induces chronical activation of this pathway. The mitochondrial nuclease ENDOG was found to influence PI3K/AKT activity in somatic cells. Our aim was to assess the value of ENDOG gene silencing to block cancer cell proliferation and to evaluate the relevance of ENDOG as prognostic marker. In vivo, in vitro and in silico experiments show that Endog/ENDOG silencing blunts proliferation of tumor cells dependent on high p-AKT/low PTEN activity and that ENDOG has prognostic value in specific cancer types.EndoG influences mitochondrial DNA replication and is involved in somatic cell proliferation. Here, we investigated the effect of ENDOG/Endog expression on proliferation in different tumor models. Noteworthy, ENDOG deficiency reduced proliferation of endometrial tumor cells expressing low PTEN/high p-AKT levels, and Endog deletion blunted the growth of PTEN-deficient 3D endometrial cultures. Furthermore, ENDOG silencing reduced proliferation of follicular thyroid carcinoma and glioblastoma cell lines with high p-AKT expression. High ENDOG expression was associated with a short time to treatment in a cohort of patients with chronic lymphocytic leukemia (CLL), a B-cell lymphoid neoplasm with activation of PI3K/AKT. This clinical impact was observed in the less aggressive CLL subtype with mutated IGHV in which high ENDOG and low PTEN levels were associated with worse outcome. In summary, our results show that reducing ENDOG expression hinders growth of some tumors characterized by low PTEN activity and high p-AKT expression and that ENDOG has prognostic value for some cancer types.

Deleterious Mutations in the TPO Gene Associated with Familial Thyroid Follicular Cell Carcinoma in Dutch German Longhaired Pointers.

Familial thyroid cancer originating from follicular cells accounts for 5–15% of all the thyroid carcinoma cases in humans. Previously, we described thyroid follicular cell carcinomas in a large number of the Dutch German longhaired pointers (GLPs) with a likely autosomal recessive inheritance pattern. Here, we investigated the genetic causes of the disease using a combined approach of genome-wide association study and runs of homozygosity (ROH) analysis based on 170k SNP array genotype data and whole-genome sequences. A region 0–5 Mb on chromosome 17 was identified to be associated with the disease. Whole-genome sequencing revealed many mutations fitting the recessive inheritance pattern in this region including two deleterious mutations in the TPO gene, chr17:800788G>A (686F>V) and chr17:805276C>T (845T>M). These two SNP were subsequently genotyped in 186 GLPs (59 affected and 127 unaffected) and confirmed to be highly associated with the disease. The recessive genotypes had higher relative risks of 16.94 and 16.64 compared to homozygous genotypes for the reference alleles, respectively. This study provides novel insight into the genetic causes leading to the familial thyroid follicular cell carcinoma, and we were able to develop a genetic test to screen susceptible dogs.

Long Noncoding RNA GAS5 Targeting miR-221-3p/Cyclin-Dependent Kinase Inhibitor 2B Axis Regulates Follicular Thyroid Carcinoma Cell Cycle and Proliferation

Introduction: Follicular thyroid carcinoma (FTC) is more aggressive than the most common papillary thyroid carcinoma (PTC). However, the current research on FTC is less than PTC. Here, we investigated the effects of long noncoding RNA (lncRNA) GAS5 and miR-221-3p in FTC. Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to detect GAS5 and miR-221-3p expression in the FTC tissues and cells. Cell proliferation was assessed by CCK8 and EdU assays. Flow cytometry was performed to determine the cell cycle. The dual-luciferase reporter assay was employed to validate the binding relationship of GAS5/miR-221-3p and miR-221-3p/cyclin-dependent kinase inhibitor 2B (CDKN2B). Western blot was conducted to measure the protein level of CDKN2B. Results: Our results displayed that GAS5 was downregulated, while miR-221-3p was upregulated in FTC tissues and cells. What’s more, overexpression of GAS5 or miR-221-3p inhibition induced G0/G1 phase arrest and inhibited cell proliferation of FTC cells. GAS5 acted as a sponge of miR-221-3p, and CDKN2B was a target gene of miR-221-3p. Additionally, GAS5 inhibited cell cycle and proliferation of FTC cells via reducing miR-221-3p expression to enhance CDKN2B expression. Conclusion: GAS5 induced G0/G1 phase arrest and inhibited cell proliferation via targeting miR-221-3p/CDKN2B axis in FTC. Thus, GAS5 may be a potential therapeutic target for the treatment of FTC.

Validation of Immunohistochemistry for Canine Proteins Involved in Thyroid Iodine Uptake and Their Expression in Canine Follicular Cell Thyroid Carcinomas (FTCs) and FTC-Derived Organoids.

Thyrotropin receptor (TSHR), sodium iodide symporter (NIS), pendrin, and thyroid peroxidase (TPO) are essential for the uptake of iodine by follicular thyroid cells. The aim of this study was to establish immunohistochemistry (IHC) protocols for TSHR, NIS, pendrin, and TPO in canine tissues and characterize their expression in organoids derived from canine follicular cell thyroid carcinoma (FTC) and in the respective primary tumors. This constitutes a fundamental step to establish organoids as a model to study the uptake of iodine in canine FTC. Commercially available antibodies directed against human proteins were selected. Antibody specificity was confirmed by western blot using lysates of the HTori-3 human thyroid cell line and healthy canine thyroid gland. IHC was validated using HTori-3 cells and a set of canine normal tissues including healthy thyroid gland. The expression of TSHR, NIS, pendrin, and TPO was evaluated in 3 organoid lines derived from FTC and respective primary tumors. All 4 antibodies produced specific bands by western blot and cytoplasmic labeling in follicular cells by IHC in both human HTori-3 cells and canine thyroid gland. NIS also showed basolateral membrane immunolabeling in follicular cells. All 4 proteins were highly expressed in organoids derived from FTC. The expression was similar or higher compared to the primary tumors. The results of this study characterize organoids derived from canine FTC as a suitable in vitro model to investigate iodine uptake, opening new research possibilities in the field of canine thyroid cancer therapy.