Thyroid Cancer Cell Proliferation Research Articles

SREBP1 as a Potential Biomarker Promotes Cell Proliferation and Invasion in Thyroid Cancer

SREBP1 as a Potential Biomarker Promotes Cell Proliferation and Invasion in Thyroid Cancer

Long non‑coding RNA small nucleolar RNA host gene 7 is upregulated and promotes cell proliferation in thyroid cancer

Thyroid cancer (THCA) is one of the most common types of endocrine cancer worldwide. However, the mechanisms underlying THCA progression have not been fully elucidated. Recent studies have demonstrated that long non-coding RNAs (lncRNAs) are dysregulated in human diseases, and are involved in regulating various biological processes. Furthermore, several reports have indicated that lncRNAs serve important roles in THCA. In the present study, a dataset from The Cancer Genome Atlas was used to analyze the expression levels and the clinical information of small nucleolar RNA host gene 7 (SNHG7) in THCA. Starbase was used to construct the competing endogenous RNA network. The Molecule Annotation System was used to analyze the data from Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases. Furthermore, cell proliferation and cell cycle assays were used to detect the functions of SNHG7 in THCA. The present study revealed for the first time, to the best of our knowledge, that SNHG7 is markedly upregulated in THCA samples following analysis of The Cancer Genome Atlas datasets. SNHG7 expression was higher in advanced stage compared with early stage THCA samples. In addition, high expression levels of SNHG7 were associated with shorter survival times in THCA patients compared with low expression levels. Bioinformatics analysis revealed that SNHG7 was associated with the processes of ‘protein translation’, ‘viral life cycle’, ‘RNA processing’, ‘mRNA splicing’, ‘histone ubiquitination’, ‘endoplasmic reticulum-to-Golgi vesicle-mediated transport’, ‘sister chromatid cohesion’, ‘DNA damage checkpoint regulation’, ‘translation’ and ‘the spliceosome’. Additionally, knockdown of SNHG7 significantly inhibited thyroid cancer cell proliferation and cell cycle progression in vitro. Taken together, the results obtained in the present study suggested that SNHG7 may serve as a novel therapeutic and prognostic target for THCA.

Impact of Antioxidant Natural Compounds on the Thyroid Gland and Implication of the Keap1/Nrf2 Signaling Pathway.

Natural compounds with potential antioxidant properties have been used in the form of food supplements or extracts with the intent to prevent or treat various diseases. Many of these compounds can activate the cytoprotective Nrf2 pathway. Besides, some of them are known to impact the thyroid gland, often with potential side-effects, but in other instances, with potential utility in the treatment of thyroid disorders. In view of recent data regarding the multiple roles of Nrf2 in the thyroid, this review summarizes the current bibliography on natural compounds that can have an effect on thyroid gland physiology and pathophysiology, and it discusses the potential implication of the Nrf2 system in the respective mechanisms. Literature searches for articles from 1950 to 2018 were performed in PubMed and Google Scholar using relevant keywords about phytochemicals, Nrf2 and thyroid. Natural substances were categorized into phenolic compounds, sulfur-containing compounds, quinones, terpenoids, or under the general category of plant extracts. For individual compounds in each category, respective data were summarized, as derived from in vitro (cell lines), preclinical (animal models) and clinical studies. The main emerging themes were as follows: phenolic compounds often showed potential to affect the production of thyroid hormones; sulfur-containing compounds impacted the pathogenesis of goiter and the proliferation of thyroid cancer cells; while quinones and terpenoids modified Nrf2 signaling in thyroid cell lines. Natural compounds that modify the activity of the Nrf2 pathway should be evaluated carefully, not only for their potential to be used as therapeutic agents for thyroid disorders, but also for their thyroidal safety when used for the prevention and treatment of non-thyroidal diseases.

Effect of bone morphogenetic protein-4 on proliferation, apoptosis and migration of thyroid cancer cells and its mechanism

Objective To investigate the effects of bone morphogenetic protein-4 (BMP-4) on the proliferation, apoptosis and migration of thyroid cancer cells through extracellular signal-regulated kinase 1/2 (ERK1/2)/p38 mitogen-activated protein kinase (p38MAPK) signaling pathway. Methods Thyroid papillary carcinoma (TPC)-1 cells (thyroid papillary carcinoma cells) were randomly divided into blank control group (BC), negative control group (NC) and interference group (KD). Cells in BC group were not transfected. NC group cells were transfected with negative control plasmids. KD group cells were transfected with the silenced BMP-4 plasmid. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to determine the BMP-4 mRNA levels. Cell counting kit-8 (CCK-8) was used to measure cell proliferation. Flow cytometry was used to determine cell apoptosis. Transwell chamber and scratch assay was used to determine cell migration capacity. The levels of ERK1/2, phosphorylated ERK1/2 (p-ERK1/2), p38MAPK and p-p38MAPK protein were determined by Western blotting. Results The level of BMP-4 mRNA in TPC-1 cells (5.86±0.42) was higher than that in HT-ori3 cells (normal thyroid cells) (1.00±0.03, t=30.537, P 0.05). Conclusion Silencing BMP-4 can inhibit the proliferation and migration of thyroid cancer cells and promote apoptosis, which may be related to down-regulation of ERK1/2/p38MAPK signaling pathway. Key words: Bone morphogenetic protein-4; Extracellular regulated protein kinase; p38 mitogen-activated protein kinase; Thyroid cancer; Proliferation; Apoptosis; Migration

Sulfasalazine suppresses thyroid cancer cell proliferation and metastasis through T-cell originated protein kinase.

Thyroid cancer patients with radioactive iodine-refractory or rapidly progressing presentation require effective treatment. T-cell originated protein kinase (TOPK) is highly expressed in a number of different tumor types, where it promotes proliferation and metastasis. However, the expression of TOPK in thyroid cancer is poorly documented. Therefore, immunohistochemistry was used to detect the expression of TOPK in thyroid cancer tissues, and its clinical significance in this disease was investigated. Sulfasalazine, a targeted inhibitor of TOPK that directly binds the protein with a dissociation constant (Kd) of 228 µM, was also investigated using microscale thermophoresis. Sulfasalazine inhibited TOPK activity, as determined by an in vitro pull-down assay. Furthermore, sulfasalazine inhibited the proliferation and metastasis of thyroid cancer cells. The results indicated that TOPK may be a potential therapeutic target and diagnostic biomarker for thyroid cancer and may be used as an index to evaluate malignant thyroid nodules. Therefore, sulfasalazine is a potential novel compound for the targeted treatment of thyroid cancer.

Expression of ANGPTL2 and its impact on papillary thyroid cancer

BackgroundAlthough the most thyroid carcinoma patients have good prognosis, around 20% of papillary thyroid carcinoma (PTC) patients have a high rate of metastasis and recurrence after routine treatment, which causes high lethality with these patients. Tumor proliferation, metastasis, and invasion are important predictors of PTC invasiveness and are key factors in cancer-related death. Angiopoietin-like 2 (ANGPTL2), a secreted protein which belongs to the angiopoietin (ANGPTL) family, was reported to be involved in the regulation of several different type of cancer cell proliferation and metastasis. However, whether ANGPTL2 plays a role in the progression of PTC, particularly in metastasis and recurrence of PTC, remains unclear. Hence, the purpose of this study was to evaluate the level of ANGPTL2 in PTC and normal thyroid, as well as para-cancerous tissue. Furthermore, the impact of ANGPTL2 on PTC cell proliferation, metastasis, recurrence and invasion was assessed to investigate the possibility whether ANGPTL2 may become a novel target for PTC therapy and cancer prognosis.Materials and methodsThe level of ANGPTL2 in PTC and para-cancerous tissue was assessed by immunohistochemistry. The biological effect of ANGPTL2 on thyroid cancer cell proliferation and metastasis was investigated by the Cell Counting Kit-8 (CCK8) assay, cell scratch test, and transwell assay. Correlations of ANGPTL2 expression levels with proliferation, migration, and metastasis of thyroid cancer were assessed with the TCGA data set and analyzed by gene set enrichment analysis. Receiver operating characteristic analysis was used to evaluate the utility of ANGPTL2 as a biomarker for prediction of thyroid cancer. Survival analysis was performed using the thyroid cancer database in K–M Plotter to detect correlations between survival time and ANGPTL2 levels.ResultsCurrent study revealed that: (1) ANGPTL2 was highly expressed in thyroid cancer in comparison with adjacent normal thyroid tissue; (2) ANGPTL2 expression was increased with thyroid tumor progression; (3) ANGPTL2 increased proliferation of thyroid cancer cells; (4) ANGPTL2 promoted migration and invasion of thyroid cancer cells; (5) high level of ANGPTL2 in thyroid cancer patients were significantly associated with a poor prognosis. The patients showed a higher metastasis and recurrence rate.ConclusionANGPTL2 promoted and enhanced proliferation, metastasis, and invasion of thyroid cancer cells. ANGPTL2 may be considered as a potential biomarker for diagnosis and prognosis of thyroid cancer patients. Further evaluation needs to be done to analyze the possibility of taking ANGPTL2 as a prognostic marker and therapeutic target for papillary thyroid cancer.

MicroRNA-592 suppresses the malignant phenotypes of thyroid cancer by regulating lncRNA NEAT1 and downregulating NOVA1.

Numerous studies have demonstrated that various microRNAs (miRs) are aberrantly expressed in thyroid cancer and play critical roles in thyroid cancer malignancy. The aberrant expression of miR‑592 has frequently been reported in multiple human cancer types; however, its expression profile and functions in thyroid cancer remain poorly understood. Reverse transcription‑quantitative polymerase chain reaction was carried out to determine the expression profile of miR‑592 in thyroid cancer tissues and cell lines. The regulatory effects of miR‑592 upregulation on thyroid cancer cell proliferation, migration, and invasion invitro, and tumor growth invivo were investigated using a CCK‑8 assay, migration and invasion assays, and a xenograft tumor model, respectively. Furthermore, the mechanisms underlying miR‑592‑mediated suppression of the aggressive phenotypes of thyroid cancer cells were explored in detail. The results indicated that miR‑592 was significantly downregulated in thyroid cancer samples, and its downregulation was associated with lymph node metastasis and tumor‑node‑metastasis stage. Patients with thyroid cancer and low miR‑592 expression exhibited shorter overall survival than patients with high miR‑592 expression. Overexpression of miR‑592 resulted in decreased cell proliferation, migration, and invasion in thyroid cancer. In addition, neuro‑oncological ventral antigen1 (NOVA1) was identified as a novel target gene of miR‑592 in thyroid cancer cells. Furthermore, ectopic NOVA1 expression may effectively abolish the tumor‑suppressing effects of miR‑592 overexpression in thyroid cancer cells. Notably, the lncRNA NEAT1 was proposed to function as a sponge of miR‑592 in thyroid cancer cells, thereby regulating NOVA1 expression. Finally, resuming miR‑592 expression significantly impaired thyroid cancer tumor growth invivo. The results indicated that the NEAT1/miR‑592/NOVA1 pathway may play regulatory roles in thyroid cancer malignancy invitro and invivo. Our findings may provide novel insight into the pathogenesis of thyroid cancer. Therefore, this pathway may be an effective target for treating patients with this disease.

Long non-coding RNA NR2F1-AS1 promoted proliferation and migration yet suppressed apoptosis of thyroid cancer cells through regulating miRNA-338-3p/CCND1 axis.

Thyroid cancer (TC) is a prevalent endocrine malignant cancer whose pathogenic mechanism remains unclear. The aim of the study was to investigate the roles of long non‐coding RNA (lncRNA) NR2F1‐AS1/miRNA‐338‐3P/CCND1 axis in TC progression. Differentially expressed lncRNAs and mRNAs in TC tissues were screened out and visualized by R program. Relative expression of NR2F1‐AS1, miRNA‐338‐3p and cyclin D1 (CCND1) was determined by quantitative real time polymerase chain reaction. In addition, Western blot analysis was adopted for evaluation of protein expression of CCND1. Targeted relationships between NR2F1‐AS1 and miRNA‐338‐3p, as well as miRNA‐338‐3p and CCND1 were predicted using bioinformatics analysis and validated by dual‐luciferase reporter gene assay. Besides, tumour xenograft assay was adopted for verification of the role of NR2F1‐AS1 in TC in vivo. NR2F1‐AS1 and CCND1 were overexpressed, whereas miRNA‐338‐3p was down‐regulated in TC tissues and cell lines. Down‐regulation of NR2F1‐AS1 and CCND1 suppressed proliferation and migration of TC cells yet greatly enhanced cell apoptotic rate. Silence of NR2F1‐AS1 significantly suppressed TC tumorigenesis in vivo. NR2F1‐AS1 sponged miRNA‐338‐3p to up‐regulate CCND1 expression to promote TC progression. Our study demonstrated that up‐regulation of NR2F1‐AS1 accelerated TC progression through regulating miRNA‐338‐3P/CCND1 axis.

Abstract 795: GDF15, the putative mitokine factor, promotes tumor progression in thyroid cancer via STAT3 regulation

Abstract Purpose: As mitochondrial dysfunction is an integral component during the initial stages of tumorigenesis, mechanistic insight on the interaction between these two pathways may lead to novel therapeutic approaches. The aim of this study is to investigate the role of GDF15, the putative mitokine factor in thyroid tumorigenesis. Methods: Expression levels of GDF15 was investigated using The Cancer Genome Atlas database. The role of GDF15 in tumor aggressiveness was investigated by analyzing the effects of knock-down of GDF15 on proliferation, migration, and invasion of thyroid cancer cells. Serum levels of GDF15 were measured in healthy subjects and patients with papillary thyroid cancer (PTC), and expression of GDF15 by IHC was investigated in PTC samples. To investigate the exact mechanism of GDF15 in thyroid cancer, both IPA analysis and GSEA analysis were performed using TCGA database and RNA seq data from Chungnam national university hospital. Results: TCGA data revealed the higher expression of GDF15 in tumor samples compared to in paired non-tumor samples. Thyroid cancer cells with knock-down of GDF15 revealed the decrease of proliferation, migration, and invasion compared to control cells via regulation of STAT3. Higher serum levels of GDF15 were found in patients with thyroid cancer than in control participants, and were significantly associated with tumor stage, lymphovascular invasion, and recurrence. In addition, patients with high GDF15 expression by IHC revealed the aggressive phenotype compared to patients with low GDF15 expression. Conclusions: GDF15 is a new biomarker for predicting tumor progression, and therapies targeting GDF15 may be effective for treating thyroid cancer. Citation Format: Yea Eun Kang, Jae Won Chang, Lihua Liu, Kyoungmin Lee, Mi Ae Lim, Seung-Nam Jung, Chan Oh, Ho-Ryun Won, Minho Shong, Bon Seok Koo. GDF15, the putative mitokine factor, promotes tumor progression in thyroid cancer via STAT3 regulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 795.

Linc00210 enhances the malignancy of thyroid cancer cells by modulating miR-195-5p/IGF1R/Akt axis.

Emerging evidence has indicated that long noncoding RNA (lncRNAs) play crucial roles in regulating thyroid cancer (TC) development. Linc00210 is a newly identified lncRNA which plays an oncogenic role in hepatocellular carcinoma and nasopharyngeal carcinoma, but whether Linc00210 can modulate the development of TC remains elusive. Here, we found that Linc00210 expression was upregulated in TC tissues compared to the matched noncancerous tissues. Overexpression of Linc00210 augmented the proliferation, migration, and invasion of TC cells. Mechanistically, Linc00210 served as a sponge for miR-195-5p, thereby counteracting its ability in downregulating the expression of IGF1R and the activation of PI3K/Akt signaling. Moreover, inhibition of Linc00210 suppressed the growth of TC cells in nude mice. Our findings for the first time uncovered the oncogenic property of Linc00210 in TC.

MicroRNA‑873 inhibits the progression of thyroid cancer by directly targeting ZEB1.

Recent evidence has revealed that certain microRNAs (miRNAs) are dysregulated in thyroid cancer (TC), which has been associated with the pathogenesis and development of TC. Therefore, improved understanding of the detailed roles of miRNAs that are aberrantly expressed in TC may aid the development of valuable therapeutic methods for the management of patients with this malignancy. In this study, miRNA‑873‑5p (miR‑873) expression in TC tissues and cell lines was detected by reverse‑transcription quantitative polymerase chain reaction (RT‑qPCR). MTT and cell invasion assays were performed to investigate the effects of miR‑873 overexpression on TC cell proliferation and invasion invitro. Bioinformatics analysis, luciferase reporter assay, RT‑qPCR and western blot analysis were employed to evaluate whether zinc finger E‑box‑binding homeobox 1 (ZEB1) is a direct target of miR‑873 in TC cells. The present study reported that miR‑873 was weakly expressed in human TC tissues and cell lines. Functionally, miR‑873 overexpression suppressed TC cell proliferation and invasion invitro. Mechanistically, ZEB1 was predicted to be a putative target of miR‑873. In addition, miR‑873 was proposed to directly bind to the 3'‑untranslated region of ZEB1 and decrease its expression in TC cells at the mRNA and protein levels. Furthermore, ZEB1 expression was significantly upregulated in TC tissues and negatively correlated with miR‑873 expression. Furthermore, ZEB1 restoration partially reversed the suppressive effects of miR‑873 overexpression on TC cell proliferation and invasion. These results demonstrated that miR‑873 may serve tumour‑suppressive roles in the progression of TC, and its suppressive effects could be mediated by the inhibition of ZEB1. Therefore, miR‑873 may be a valuable therapeutic target in the management of patients with TC.

Knockdown of long noncoding RNA SNHG7 inhibits the proliferation and promotes apoptosis of thyroid cancer cells by downregulating BDNF.

Recently, long noncoding RNAs (lncRNAs) have got much attention for their role in tumor progression. LncRNA small nucleolar RNA host gene 7 (SNHG7) was studied in this research to identify how it affects the development of thyroid cancer (TC). SNHG7 expression was detected by quantitative Real-time polymerase chain reaction (qRT-PCR) in both TC cells and tissue samples. Pearson's Chi-square test was used to determine the association of SNHG7 expression with several clinicopathological factors. Moreover, colony formation assay, cell proliferation and cell apoptosis assay were conducted. In addition, by performing qRT-PCR and Western blot assay, the underlying mechanism was explored. SNHG7 expression level was higher in TC samples than that in corresponding ones. The SNHG7 expression was associated with tumor size and TNM stage. Moreover, TC cell proliferation was inhibited, and TC cell apoptosis was induced after SNHG7 was knocked down in vitro. Moreover, the mRNA and protein expressions of brain-derived neurotrophic factor (BDNF) were downregulated after knockdown of SNHG7. Furthermore, the expression level of BDNF was positively related to the expression of SNHG7 in TC tissues. These results suggested that knockdown of SNHG7 could inhibit TC cell proliferation and induce cell apoptosis via downregulating BDNF, which might be a potential therapeutic target in TC.

TFAP2B overexpression contributes to tumor growth and progression of thyroid cancer through the COX-2 signaling pathway

Thyroid cancer is commonly seen in the clinic with a rapidly increasing incidence globally. COX-2 overexpression correlates with the pathologic type of thyroid carcinoma, and it has been suggested that COX-2 overexpression is associated with a poor prognosis. However, little is known about its upstream regulatory mechanism. Bioinformatics suggested that transcription factor AP-2 beta (TFAP2B) might specifically bind to the COX-2 promoter, which was confirmed by biotin-labeled COX-2 promoter pulldown and luciferase reporter assays. We performed western blot and immunohistochemical staining to detect the expression of TFAP2B/COX-2 in thyroid cancer tissues (T) and the matched adjacent noncarcinoma tissues (ANT), and investigated the relationship between TFAP2B/COX-2 expression and clinical pathological factors in thyroid cancer patients. Afterward, MTS, colony formation, cell-apoptosis assay, transwell-invasion and scratch assays were performed to examine the proliferation, apoptosis, invasion, and migration of thyroid cancer cells with TFAP2B knocked down or overexpressed. The mouse xenograft experiment was performed to study in vivo the proliferation of thyroid cancer cells with TFAP2B knocked down or overexpressed. We found that TFAP2B bound to the promoter of COX-2 to activate its expression. Western blot and immunohistochemistry showed that TFAP2B/COX-2 was highly expressed in thyroid cancer, and high TFAP2B and COX-2 expression was associated with aggressive clinicopathological features in thyroid cancer. TFAP2B mediated thyroid cancer cell proliferation, apoptosis, invasion, and migration via the COX-2 signaling pathway in vitro and in vivo. TFAP2B bound to the promoter of COX-2 to activate its expression, indicating that TFAP2B is a critical regulatory molecule in the COX-2 signaling pathway that promoted tumor progression in thyroid cancer.

MicroRNA-30a suppresses papillary thyroid cancer cell proliferation, migration and invasion by directly targeting E2F7.

microRNA (miRNA/miR)-30a, a tumor-associated miRNA, has been implicated in the tumorigenesis and progression of different types of human cancer. Thyroid cancer is a common endocrine malignancy, of which papillary thyroid cancer (PTC) accounts for ~80–90% of all TC. However, the effect of miR-30a in PTC is yet to be fully elucidated. The TPC-1 human PTC cell line, as well as the normal human thyroid cell line (HT-ori3), were utilized in the current study. The PTC cell line was transfected with a miR-30a mimic. Subsequently, reverse transcription-quantitative polymerase chain reaction was performed to detect the expression of miR-30a and E2F transcription factor 7 (E2F7). Cell proliferation was assessed via a MTT assay and transwell migration and invasion assays were performed to detect the migration and invasion of PTC cells. A dual-luciferase reporter assay was also utilized to clarify the association between miR-30a and E2F7. The results of the current study revealed that miR-30a was significantly downregulated in TPC-1 cells compared with HT-ori3 cells and that the expression of E2F7 was significantly upregulated in PTC cells. The upregulation of miR-30a also inhibited the proliferation, migration and invasion of PTC cells. Furthermore, the luciferase assay revealed that miR-30a binds to the 3′-UTR of E2F7. Additionally, the overexpression of miR-30a decreased E2F7 levels in TPC-1 cells. These results indicate that miR-30a functions as a tumor suppressor in PTC by direct targeting E2F7 and that miR-30a may be a novel therapeutic target for patients with PTC.

LncRNA MCM3AP-AS1 promotes proliferation and invasion through regulating miR-211-5p/SPARC axis in papillary thyroid cancer.

Long non-coding RNAs (lncRNAs) are an emerging class of regulators in cancer. A lncRNA, MCM3AP-AS1, has been demonstrated as a versatile mediator in many cancers, except papillary thyroid cancer. The aim of this study is to investigate the role and mechanism of MCM3AP-AS1 in papillary thyroid cancer. Quantitative real-time PCR was used to assess the level of MCM3AP-AS1 and miR-211-5p in papillary thyroid cancer tissues and cells. Western blot was used to detect E-cadherin and secreted protein acidic and cysteine rich (SPARC) protein levels. CCK-8, scratch wound assay, and transwell assay were used to evaluate papillary thyroid cancer cell proliferation, migration, and invasion, respectively. BLAST alignment and luciferase assay were used to explore the interaction among MCM3AP-AS1, mi/r-211, and SPARC. In papillary thyroid cancer, MCM3AP-AS1 was upregulated, while miR-211 was downregulated. MCM3AP-AS1 overexpression promoted papillary thyroid cancer proliferation, migration, and invasion. Further, MCM3AP-AS1 was shown to be negatively correlated with miR-211-5p. We next validated that miR-211-5p overexpression could reverse the promoting role of MCM3AP-AS1 in papillary thyroid cancer, whereby SPARC plays an important regulating role. In vivo, we confirmed the anti-tumor role of MCM3AP-AS1 silencing and the close relation among MCM3AP-AS1, miR-211-5p, and SPARC. MCM3AP-AS1 promotes papillary thyroid cancer by regulating the MCM3AP-AS1/miR-211-5p/SPARC axis, which could potentially be a therapeutic target in papillary thyroid cancer.

NECTIN4 promotes papillary thyroid cancer cell proliferation, migration, and invasion and triggers EMT by activating AKT.

Papillary thyroid cancer (PTC) is the most frequent type of malignant thyroid cancer, but its molecular mechanisms remain unknown. To better understand the tumorigenesis and progression of PTC, we conducted a comprehensive analysis of the whole-transcriptome resequencing of paired PTC and normal thyroid tissues. Nectin cell adhesion molecule 4 (NECTIN4) was significantly overexpressed in thyroid carcinoma compared with that in matched normal tissue. We also assessed the relation between the expression level of NECTIN4 and the clinicopathological features of PTC in The Cancer Genome Atlas database, and results showed that upregulated NECTIN4 is associated with lymph node metastasis (P<0.001) and tumor size (P=0.017). The biological function of NECTIN4 was also investigated by using the PTC cell lines TPC-1 and KTC-1. In vitro experiments demonstrated that NECTIN4 downregulation significantly inhibits the colony formation, proliferation, migration, and invasion of PTC cell lines. NECTIN4 could modulate the expression of epithelial–mesenchymal transition-related proteins via the PI3K/AKT pathway, and SC79, an AKT phosphorylation activator, could reverse the si-RNA knockdown effect. In addition, after the use of AKT inhibitors (LY 294,002), we found that SiRNA have similar effect with AKT inhibitors. Taking the results together, the current study shows that NECTIN4 has important biological implications in the tumorigenesis and metastasis of PTC and may be a potential therapeutic target for the disease.

Junctional adhesion molecule-A is down-regulated in anaplastic thyroid carcinomas and reduces cancer cell aggressiveness by modulating p53 and GSK3 α/β pathways.

Junctional adhesion molecule A (JAM-A) is a transmembrane protein that contributes to different biological process, including the epithelial to mesenchymal transition (EMT). Through an EMT profiler array, we explored the molecular players associated with human thyroid cancer progression and identified JAM-A as one of the genes mostly deregulated. The quantitative real-time polymerase chain reaction and immunohistochemistry analyses showed that downregulation of JAM-A occurred in anaplastic thyroid carcinoma (ATC) compared with normal thyroid (NT) and papillary thyroid carcinoma (PTC) tissues and correlated with extrathyroid infiltration, tumor size, and ATC histotype. In ATC cell lines, JAM-A restoration suppressed malignant hallmarks of transformation including cell proliferation, motility, and transendothelial migration. Accordingly, knockdown of JAM-A enhanced thyroid cancer cell proliferation and motility in PTC cells. Through the proteome profiler human phospho-kinase array, we demonstrated that higher expression of JAM-A was associated with a significant increased level of phosphorylation of p53 and GSK3 α/β proteins. In conclusion, our findings highlight a novel role of JAM-A in thyroid cancer progression and suggest that JAM-A restoration could have potential clinical relevance in thyroid cancer treatment.

SIRT4 inhibits the proliferation, migration, and invasion abilities of thyroid cancer cells by inhibiting glutamine metabolism.

BackgroundSIRT4, a protein localized in the mitochondria, is one of the least characteristic members of the sirtuin family. It is known that SIRT4 has deacetylase activity and plays a role in energy metabolism, but little is known about its possible role in carcinogenesis. Recently, several studies have suggested that SIRT4 may function as either a tumor oncogene or a tumor suppressor gene. However, its relationship with thyroid cancer remains unclear.MethodsWe stably overexpressed SIRT4 or silenced its expression in the human thyroid cancer cell line BCPAP by means of lentiviral vectors. We conducted a variety of tests, such as CCK-8, wound healing, migration, and invasion assays, to investigate the role of SIRT4 in the proliferation, migration, and invasion abilities of thyroid cancer cells. We also investigated the effects of SIRT4 overexpression on cell cycle progression and apoptosis of BCPAP cells and studied the role of glutamine metabolism in the effects of SIRT4 on BCPAP cell migration and invasion. Finally, we analyzed SIRT4 expression levels in thyroid cancer specimens by immunohistochemistry and investigated their association with clinicopathological features.ResultsOverexpression of SIRT4 inhibited the proliferation, migration, and invasion abilities of BCPAP thyroid cancer cells, blocked the cell cycle in the G0/G1 phase, and induced apoptosis. Mechanistically, SIRT4 inhibited BCPAP migration and invasion by inhibiting glutamine metabolism. Moreover, we found that SIRT4 protein levels in thyroid cancer tissues were markedly lower than in their non-neoplastic tissue counterparts (P<0.001).ConclusionSIRT4 plays a pivotal role in the growth and metastasis of thyroid cancer cells and could be a potential therapeutic target in thyroid cancer.

MicroRNA-510-5p promotes thyroid cancer cell proliferation, migration, and invasion through suppressing SNHG15.

SNHG15 has been suggested to be correlated with clinical progression and prognosis, and function as tumor suppressive long noncoding RNA in thyroid cancer at our previous study. SNHG15 was proposed to be a potential target for miR-510-5p at LncBase Predicted database. Thus, the aim of this study was to explore the relationship between miR-510-5p and SNHG15 in thyroid cancer, and the clinical significance of miR-510-5p in patients with thyroid cancer. In our results, levels of miR-510-5p expression were increased in thyroid cancer tissues and cell lines compared with adjacent normal thyroid tissues and normal thyroid cell line, respectively. There was a statistically negative correlation between SNHG15 expression and miR-510-5p expression in thyroid cancer tissues. Moreover, miR-510-5p directly bound to SNHG15, and negatively regulated SNHG15 expression in thyroid cancer cells. Furthermore, miR-510-5p promoted thyroid cancer cell proliferation, migration, and invasion through suppressing SNHG15. Finally, high miR-510-5p expression was observed in tumor tissues with advanced clinical stage or lymph node metastasis. In conclusion, we provide evidence to support a pivotal role for miR-510-5p in regulating thyroid cancer cell proliferation, migration, and invasion.

Upregulated circular RNA circ_0025033 promotes papillary thyroid cancer cell proliferation and invasion via sponging miR-1231 and miR-1304

Recent evidence revealed that circular RNAs (circRNAs) are key regulators for tumorigenesis. However, their roles in papillary thyroid cancer (PTC) is not fully understood. In the current work, the differentially expressed circRNAs between PTC tissues and adjacent noncancerous tissue samples were screened by circRNA microarray. We further selected the highest expressed circRNA (circ_0025033) in tumorous tissues for the study. qRT-PCR was used to measure the level of circ_0025033 in PTC tissue samples and cell lines. Gain/loss of function assays were carried out to determine the effect of circ_0025033 on cell viability, clone-forming, apoptosis, migration and invasion. Dual-luciferase reporter assays were conducted to investigate the mechanisms of circ_0025033 in PTC. The data showed that circ_0025033 aggravated cell proliferation, migration and invasion and inhibited cell apoptosis. Additionally, circ_0025033 could directly sponge miR-1231 and miR-1304 in PTC cells. Furthermore, the oncogenic role of circ_0025033 is dependent on its suppression of miR-1231 and miR-1304. Collectively, this work uncovers a novel signal of circ_0025033/miR-1231/miR-1304 involved in PTC initiation and progression.