Thyroid Carcinoma Cell Lines Research Articles

Long Non-Coding RNA LUCAT1 Promotes Progression of Thyroid Carcinoma by Reinforcing ADAM10 Expression Through Sequestering microRNA-493.

BackgroundLong non-coding RNA (lncRNA) LUCAT1 has recently been recognized as an oncogene in several malignancies. This study was launched to probe its role in thyroid carcinoma (TC) development and the implicated molecules.MethodsLUCAT1 expression in TC cell lines and in normal thyroid follicular epithelial cell line Nthy-ori3-1 was determined by RT-qPCR. Binding relationships between LUCAT1 and microRNA (miR)-493, and between miR-493 and a disintegrin and metalloproteinase-10 (ADAM10) were predicted on a bioinformatics system and then validated through luciferase reporter gene assays. Expression of miR-493 and ADAM10 in TC cells was determined. Gain- and loss-of functions of LUCAT1, miR-493 and ADAM10 were performed to explore their influences on the behaviors of TC cells. Xenograft tumors were induced in nude mice for in vivo studies.ResultsLUCAT1 and ADAM10 were highly expressed, while miR-493 was poorly expressed in TC cell lines. LUCAT1 served as a miR-493 sponge to upregulate ADAM10 expression. Silencing of LUCAT1 discouraged proliferation, invasion, and migration but triggered apoptosis of TC cells. By contrast, these changes were abrogated by further miR-493 inhibition or ADAM10 upregulation. The in vitro experiment results were reproduced in vivo. In addition, miR-493 inhibition or ADAM10 overexpression was found to increase the phosphorylation of STAT3 in cells.ConclusionThis study evidenced that LUCAT1 increases ADAM10 expression through sequestering miR-493, leading to JAK-STAT activation and TC cell growth and metastasis. LUCAT1 and ADAM10 may serve as therapeutic targets for TC treatment.

Beta-elemene inhibits differentiated thyroid carcinoma metastasis by reducing cellular proliferation, metabolism and invasion ability.

BackgroundAccelerated glycolysis is a characteristic of carcinoma. The herb-derived compound, beta (β)-elemene, has shown promising anticancer effects against various tumors by inhibiting aerobic glycolysis. However, its activity against thyroid carcinoma and the mechanism is still unknown.MethodsDifferentiated thyroid carcinoma (DTC) cell lines, including papillary thyroid carcinoma (PTC) cell lines (IHH-4, TPC-1, K1), and follicular thyroid carcinoma (FTC) cell line (FTC133) were treated with different concentration of β-elemene. The viability of DTC cells was analyzed using the CCK8 method. Cell cycle and apoptosis analysis were performed by flow cytometry and western blotting. The cell invasion ability was evaluated in Transwell assays. Energy metabolism in living cells was measured using a Seahorse XF analyzer. The antitumor effects of β-elemene were analyzed in vivo in a nude mouse xenograft tumors model.ResultsCCK8 assays showed β-elemene significantly inhibited DTC cell proliferation in a dose- and time-dependent manner. β-elemene promoted cell apoptosis, with increased expression of cleaved caspase-9 and decreased BCL-2 expression. Transwell assays showed that β-elemene significantly inhibited the invasion ability of DTC cells. β-elemene also reduced angiogenesis by decreasing VEGF expression in DTC cells. β-elemene reduces the basal oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and maximal glycolytic capacity as well as maximal respiration and ATP production. Moreover, β-elemene inhibited tumor growth in a mouse xenograft model in vivo.ConclusionsIn this study, we have provided the first evidence of the antitumor effects of β-elemene, which was shown to inhibit cell proliferation, promote apoptosis, induce cell cycle arrest, inhibit cell invasion ability and reduce angiogenesis. Furthermore, we showed that β-elemene significantly inhibits the respiratory and glycolytic ability of human DTC cells. Thus, our findings show the potential of β-elemene as a novel treatment for DTC.

Circ_0000144 facilitates the progression of thyroid cancer via the miR‐217/AKT3 pathway

Thyroid carcinoma (TC) is the most common malignancy of the endocrine system. Circular RNA (circRNA) is vital in the regulation of tumor progression. Circ_0000144 serves as a novel oncogenic circRNA, and miR-217 is reported to inhibit the malignant phenotypes of cancer cells by targeting AKT3 in TC. The present study aimed to explore the regulatory mechanism of circ_0000144 and miR-217 in the progression of TC. Circ_0000144 expression in 32 pairs of TC tissues and different TC cell lines (including BCPAP, K1, H7H83, and TPC-1) was detected by employing a quantitative real-time polymerase chain reaction (qRT-PCR). Circ_0000144 small interfering RNA was used to establish loss-of-function models. Cell counting kit-8 (CCK-8), BrdU (5-bromo-2'-deoxyuridine) and transwell assays were utilized to verify the effects of circ_0000144 on TC cell proliferation, migration and invasion, respectively. Bioinformatics, western blotting, a luciferase reporter experiment and qRT-PCR were employed to confirm the relationships among circ_0000144, miR-217 and AKT3. Circ_0000144 expression was remarkably elevated in TC tissues (p<0.001) and TC cell lines. The elevation of circ_0000144 expression was markedly linked to tumor size (p=0.015), TNM stage (p=0.025) and lymph node metastasis (p=0.017) of the patients. Functional studies showed that knocking down circ_0000144 repressed the malignancy of TC cells. Furthermore, miR-217 was identified as a downstream target of circ_0000144; inhibition of miR-217 could reverse the effects induced by circ_0000144 knockdown. Moreover, circ_0000144 could regulate AKT3 expression by suppressing miR-217 expression. Circ_0000144 exerts a cancer-promoting effect on TC cells via the miR-217/AKT3 pathway.

Hsa-miR-195-5p inhibits cell proliferation of human thyroid carcinoma cells via modulation of p21/cyclin D1 axis.

BackgroundBased on existing evidence, microRNAs (miRs) are gene regulators that undertake key functions in the oncogenesis and tumor progression of every single human malignant disease, such as thyroid carcinoma (TC). Previous clinical findings showed that expression of miR-195 is down-regulated in TC, which implies that miR-195 may be practically involved in TC pathogenesis. Nevertheless, the function of hsa-miR-195-5p in TC is still largely unclear. Herein, we detected the conceivable involvement of hsa-miR-195-5p in TC cell proliferation.MethodsReal time PCR examination was performed to assess the expression level of hsa-miR-195-5p in TC cell lines TPC-1 and B-CPAP. TPC-1 cells were transfected with either hsa-miR-195-5p mimics or hsa-miR-195-5p inhibitor. After confirmation of transfection efficiency, the effect of hsa-miR-195-5p on proliferation and cell cycle of TPC-1 cells was assessed. The expression of cyclin D1 and p21 was simultaneously detected by western blotting. Moreover, targetScan 6.2 was used to predict hsa-miR-195-5p target genes. Subsequently, luciferase reporter was performed to examine whether there is a possible binding of hsa-miR-195-5p to 3’-UTR of cyclin D1 mRNA. Furthermore, cyclin D1 mRNA and protein levels were measured to check whether hsa-miR-195-5p exerts its function at the post-transcriptional level. In addition, to explore the function of cyclin D1 in TPC-1 cells overexpressing hsa-miR-195-5p, cyclin D1 siRNA was used to silence the expression of cyclin D1 in TPC-1 cells overexpressing hsa-miR-195-5p.ResultsWe quantified the expression of hsa-miR-195-5p in TC cells and normal thyroid cells and found a remarkable decrease in hsa-miR-195-5p expression in TC cells. Over-expression of hsa-miR-195-5p obviously resulted in downgraded proliferation of TC cells. Moreover, hsa-miR-195-5p caused cell arrest at the GO/G1 phase. Further in silico analyses and the dual-luciferase reporter assay confirmed that 3’-UTR of cyclin D1 is a direct target of hsa-miR-195-5p. Western blot analysis uncovered that hsa-miR-195-5p over-expression led to decreased levels of cyclin D1 and p21. In mechanistic analyses, we found that silencing of cyclin D1 reversed the inhibitory effect of hsa-miR-195-5p on the proliferation of TC cells, which indicates that hsa-miR-195-5p suppresses TC cell proliferation by adversely regulating cyclin D1.ConclusionsWe concluded that hsa-miR-195-5p is a candidate tumor-suppressor miRNA in TC and that the hsa-miR-195-5p/p21/cyclin D1 pathway could be a potential therapeutic target for TC.

SOX18 exerts tumor-suppressive functions in papillary thyroid carcinoma through inhibition of Wnt/β-catenin signaling

Sex-determining region on the Y chromosome-related high mobility group box 18 (SOX18) has emerged as a key tumor-related protein in a wide range of human tumors. Yet, the involvement of SOX18 in papillary thyroid carcinoma has not been determined. This study aimed to explore the expression and biological function of SOX18 in papillary thyroid carcinoma. There was a significant decrease in SOX18 expression in papillary thyroid carcinoma tissues compared with that in normal tissues. Low expression of SOX18 was also detected in papillary thyroid carcinoma cell lines and upregulation of SOX18 effectively repressed the proliferative, colony-forming and invasive abilities of papillary thyroid carcinoma cells in vitro. In contrast, knockdown of SOX18 in papillary thyroid carcinoma cells was associated with a significant increase in cell proliferation and invasion. Further studies revealed that SOX18 upregulation was associated with the reduced nuclear accumulation of β-catenin and the downregulation of Wnt/β-catenin signaling in thyroid carcinoma cells. Moreover, inhibition of Wnt/β-catenin signaling markedly attenuated SOX18 knockdown-evoked oncogenic effects in papillary thyroid carcinoma cells. In addition, SOX18 overexpression remarkably retarded the tumor growth of papillary thyroid carcinoma cell-derived xenograft tumors in nude mice. Taken together, these results demonstrate that SOX18 suppresses the proliferation and invasion of papillary thyroid carcinoma by inhibiting Wnt/β-catenin signaling. Our study reveals a tumor-suppressive role of SOX18 in papillary thyroid carcinoma and suggests that SOX18 is an attractive candidate target for treatment of papillary thyroid carcinoma.

MicroRNA-124-3p inhibits tumourigenesis by targeting mitogen-activated protein kinase 4 in papillary thyroid carcinoma.

The study aimed to investigate the role of miR-124-3p and its potential molecular mechanism in papillary thyroid cancer (PTC). The expression of miR-124-3p and mitogen-activated protein kinase 4 (MAP2K4) in human thyroid follicular epithelial cell line (NTHY-ORI3-1) and human papillary thyroid carcinoma cell lines (SW1736, BCPAP, TPC-1 and K1) was measured by RT-qPCR. Cell proliferation was measured by CCK-8, while cell cycle and apoptosis rate were measured by flow cytometry. Invasive ability and migrative ability were measured by transwell assay and wound healing assay, respectively. Western blot was used to detect the levels of relative proteins. In vivo, TPC-1 cells transfected with miR-124-3p mimic were subcutaneously injected into the flank of the mice to form tumour. After successful modelling, mice were divided into two groups (n = 10): Control group and miR-124-3p mimic group. The present study showed that miR-124-3p was lowly expressed, while MAP2K4 was highly expressed in PTC cell lines. Besides, miR-124-3p targeted MAP2K4 and negatively regulated MAP2K4 in TPC-1 cells. In addition, miR-124-3p inhibited the proliferation and motility, and induced apoptosis and cell cycle arrest of TPC-1 cells by inactivating MAP2K4/JNK/JunD pathway. Furthermore, miR-124-3p inhibited tumour formation by downregulating MAP2K4 level in vivo. In conclusion, the study provided a novel molecular mechanism of miR-124-3p in the progress of PTC. SIGNIFICANCE OF THE STUDY: Papillary thyroid cancer (PTC) is the most important pathological type of thyroid cancer, accounting for 80% of thyroid cancer. miR-124-3p exhibited significant inhibitory role in the transformation and development of malignant tumours. However, in PTC, the roles and its potential molecular mechanism are unclear. Here, the study investigated the roles of miR-124-3p in the progress of PTC and its potential molecular mechanism. We found that miR-124-3p inhibited the proliferation and motility, and induced apoptosis and cell cycle arrest in PTC cells. This study provided a novel molecular mechanism of miR-124-3p in the progress of PTC.

Down-Regulation of the Mammalian Target of Rapamycin (mTOR) Pathway Mediates the Effects of the Paeonol-Platinum(II) Complex in Human Thyroid Carcinoma Cells and Mouse SW1736 Tumor Xenografts.

BackgroundThis study aimed to investigate the effects of the paeonol-platinum(II) (PL-Pt[II]) complex on SW1736 human anaplastic thyroid carcinoma cell line and the BHP7-13 human thyroid papillary carcinoma cell line in vitro and on mouse SW1736 tumor xenografts in vivo.Material/MethodsThe cytotoxic effects of the PL-Pt(II) complex on SW1736 cells and BHP7-13 cells was measured using the MTT assay. Western blot measured the expression levels of cyclins, cell apoptotic proteins, and signaling proteins. DNA content and apoptosis were detected by flow cytometry. SW1736 cell thyroid tumor xenografts were established in mice followed by treatment with the PL-Pt(II) complex.ResultsTreatment of the SW1736 and BHP7-13 cells with the PL-Pt(II) complex reduced cell proliferation in a dose-dependent manner, with an IC50 of 1.25 μM and 1.0 μM, respectively, and increased the cell fraction in G0/G1phase, inhibited p53, cyclin D1, promoted p27 and p21 expression, and significantly increased the sub-G1 fraction. Treatment with the PL-Pt(II) complex increased caspase-3 degradation, reduced the expression of p-4EBP1, p-4E-BP1 and p-S6, and reduced the expression of p-ERK1/2 and p-AKT. Treatment with the PL-Pt(II) complex reduced the volume of the SW1736 mouse tumor xenografts on day 14 and day 21, and reduced AKT phosphorylation and S6 protein expression and increased degradation of caspase-3.ConclusionsThe cytotoxic effects of the PL-Pt(II) complex in human thyroid carcinoma cells, including activation of apoptosis and an increased sub-G1 cell fraction of the cell cycle, were mediated by down-regulation of the mTOR pathway.

Thyroid-related hormones as potential markers of hypoxia/ischemia

This study aimed to investigate the usefulness of the thyroid-related hormones as markers of acute systemic hypoxia/ischemia to identify deaths caused by asphyxiation due to neck compression in human autopsy cases. The following deaths from pathophysiological conditions were examined: mechanical asphyxia and acute/subacute blunt head injury; acute/subacute non-head blunt injury; sharp instrument injury as the hemorrhagic shock condition; drowning as alveolar injury; burn; and death due to cardiac dysfunction. Blood samples were collected from the left and right cardiac chambers and iliac veins, and serum triiodothyronine (T3), thyroxine (T4), thyroglobulin (Tg), and thyroid-stimulating hormone (TSH) levels were measured using electrochemiluminescence immunoassays. Two types of thyroid cell lines were used to confirm independent thyroid function under the condition of hypoxia (3% O2). The human thyroid carcinoma cell line (HOTHC) cell line derived from human anaplastic thyroid carcinoma and the UD-PTC (sample of the second resection papillary thyroid carcinoma) cell line derived from human thyroid papillary adenoma, which forms Tg retention follicles, were used to examine the secretion levels of T3, T4, and Tg hormones. The results showed a strong correlation between T3 and T4 levels in all blood sampling sites, while the TSH and Tg levels were not correlated with the other markers. Serum T3 and T4 levels were higher in cases of mechanical asphyxia and acute/subacute blunt head injury, representing hypoxic and ischemic conditions of the brain as compared to those in other causes of death. In the thyroid gland cell line, T4, T3, and Tg levels were stimulated after exposure to hypoxia for 10–30 min. These findings suggest that systemic advanced hypoxia/ischemia may cause a rapid and TSH-independent release of T3 and T4 thyroid hormones in autopsy cases. These findings demonstrate that increased thyroid-related hormone (T3 and T4) levels in the pathophysiological field may indicate systemic hypoxia/ischemia.

Circular RNA circ_0103552 promotes the invasion and migration of thyroid carcinoma cells by sponging miR-127.

The aim of this study was to examine the regulatory role of circ-0103552 in the procession of thyroid carcinoma (TC) and the related underlying mechanisms. The tissues were obtained from 56 patients diagnosed with TC in our hospital. Nthy-ori3-1 cell line and TC cell lines (TPC-1, SW579, 8505C) were purchased from American Type Culture Collection (ATCC). Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was conducted to analyze the expression of circ-0103552 in TC tissues and cell lines. Inhibition of circ-0103552 was achieved by circ-0103552 siRNA. Dual-Luciferase report assay was performed to confirm the potential target miRNA of circ-0103552. The transwell assay and wound-healing assay were designed to examine the invasion and migration ability of TC cells, respectively. Circ-0103552 was detected to be upregulated in TC tissues, as well as in TC cell lines, including TPC-1, SW579, and 8505C. The knockdown of circ-0103552 significantly reduced the invasion and migration ability in TC cells. It was predicted using the circular RNA database that microRNA-127 (miR-127) was a target miRNA of circ-0103552, which was confirmed by the Dual-Luciferase assay. Further studies revealed that circ-0103552 was involved in the invasion and migration of TC by sponging miR-127. The present study demonstrated that circ-0103552 acts as a regulator in the invasion and migration of TC by sponging miR-127.

Knockdown of Thymidine Kinase 1 Suppresses Cell Proliferation, Invasion, Migration, and Epithelial-Mesenchymal Transition in Thyroid Carcinoma Cells.

Patients with advanced thyroid carcinoma have poor prognosis with low overall survival. Unfortunately, the underlying mechanisms of thyroid carcinoma progression remain unclear. The elevated expression of thymidine kinase 1 (TK1) has been implicated in the progression of thyroid carcinoma, while the role of TK1 in thyroid carcinoma progression has not been explored. The present study aimed to determine the role TK1 in the progression of thyroid cancer and to explore the underlying molecular mechanisms. In this study, it was found that serum TK1 levels were markedly increased in the patients with thyroid nodules. Further online data mining showed that TK1 expression was upregulated in thyroid carcinoma tissues, and higher expression of TK1 was correlated with shorter disease-free survival of patients with thyroid carcinoma. Silencing of TK1 suppressed cell proliferation, invasion, migration, and epithelial–mesenchymal transition, and also induced cell apoptosis in the thyroid carcinoma cell lines. Animal studies showed that TK1 knockdown inhibited in vivo tumor growth of thyroid carcinoma cells. Importantly, miR-34a-5p was found to be downregulated in the thyroid carcinoma cells. Furthermore, miR-34a-5p targeted the 3′ untranslated region of TK1 and suppressed the expression of TK1 in thyroid carcinoma cell lines. In summary, first, these results demonstrated the upregulation of TK1 in thyroid nodules and thyroid carcinoma tissues; second, TK1 promoted thyroid carcinoma cell proliferation, invasion, and migration; lastly, TK1 was negatively regulated by miR-34a-5p. Our study may provide novel insights into the role of TK1 in regulating thyroid carcinoma progression.

1,25-Dihydroxy vitamin D3 inhibits the Ras-MEK-ERK pathway and regulates proliferation and apoptosis of papillary thyroid carcinoma

ObjectiveTo explore the effects of 1,25-dihydroxy vitamin D3 [1,25-(OH)2D3] on the proliferation and apoptosis of papillary thyroid carcinoma and to investigate its possible mechanism. Materials and methodsThe papillary thyroid carcinoma cell line TPC-1 was cultured, and the cells were divided into control group, the 1,25-(OH)2D3 group, and the 1,25-(OH)2D3 + ML-098 (Ras agonist) group. Cell proliferation was observed by MTT. The colony formation viability of cells was detected by the plate cloning assay. Cell migration was observed by the scratch assay. Apoptosis was detected by flow cytometry. The expression of Ki67 and Caspase-3, and the activity of Ras-MEK-ERK pathway were detected by western blot. ResultsCompared with the Control group, the proliferation, colony formation and migration ability of cells in the drug group were significantly decreased. The number of apoptotic cells was significantly increased, the expression of Ki67 protein was decreased, and the expression of Caspase-3 protein was upregulated. The phosphorylation levels of Ras, p-ERK1/2, and p-MEK were decreased. Compared with the drug group, the cloning and migration biological activity of cells in the 1,25-(OH)2D3 + ML-098 group was significantly enhanced (p < 0.05). The number of apoptotic cells was decreased, while the Ki67 protein level was increased. In addition, the Caspase-3 protein level was decreased, and the Ras-MEK-ERK level was also enhanced. Furthermore, the antitumor activity of 1,25-(OH)2D3 was reversed by the Ras agonist ML-098. Conclusion1,25-(OH)2D3 can inhibit the activity and promote apoptosis of the papillary thyroid carcinoma cell line TPC-1, and its mechanism may be related to the inhibition of the Ras-MEK-ERK pathway activity, thus affecting the proliferation and expression of apoptosis-related proteins.

Silencing NEAT1 suppresses thyroid carcinoma via miR-126/NEAT1/VEGFA axis.

The incidence of papillary thyroid carcinoma (PTC) has steadily increased over the recent years, making this cancer a common malignant tumor world-wide. We tested the hypothesis that Nuclear Enriched Abundant sh-NEAT1 knock-down ranscript 1 (NEAT1) is involved in the pathogenesis of PTC in vitro and in vivo. We show that NEAT1 is highly expressed in Papillary Thyroid Carcinoma cell line (PTC-1) and anaplastic thyroid cancer cell line (SW1736) as compared with the human thyroid follicular epithelial cell line (Nthy-ori 3-1). shRNA knockdown of NEAT1 led to the inhibition of cell growth, invasion, migration and Epithelial to Mesenchymal Transition (EMT) of thyroid cancer cells. This treatment increased the rate of apoptosis in SW1736 cells. Silencing of NEAT1 increased the level of its regulator, miRNA-126 and down-regulated VEGFA that sets the density of tumor vasculature. Administrtation of sh-NEAT1 also inhibited tumor growth in vivo, increased the miRNA-126 level and down-regulated VEGFA. Taken together, these results indicate that silencing NEAT1 suppresses thyroid carcinoma via miR-126/NEAT1/VEGFA axis.

C6-ceramide Inhibits the Motility of Anaplastic Thyroid Carcinoma Cells.

Anaplastic thyroid carcinoma (ATC) is an aggressive type of thyroid cancer, and its metastasis requires cell motility. Ceramide is involved in a variety of biological processes, including inflammation, cell signaling, cell motility, and induction of apoptosis, however has not previously been reported to inhibit the motility of ATC cells. We evaluated the effect of short chain C6-ceramide on motility of ATC cells. Cell motility of 8305C thyroid carcinoma cell line treated with C6-ceramide was assessed using a transwell migration assay and a pseudopodia formation assay. Treatment with 10 µM C6-ceramide resulted in significantly fewer migratory cells than control treatment in a transwell migration assay (P < 0.002). In condition medium, 82.6% of C6-ceramide-treated cells formed lamellipodia. Importantly, treatment with 10 µM C6-ceramide drastically decreased the number of cells forming lamellipodia by 17.6% (P < 0.01). Our results suggest that treatment with a low concentration of ceramide may prevent metastasis and recurrence of ATC by inhibiting cell motility. Further studies are necessary to investigate the mechanism of inhibition of cell motility by ceramide. Ceramide shows promise as a therapeutic treatment for ATC.

Effect of miR‑449a‑mediated Notch signaling pathway on the proliferation, apoptosis and invasion of papillary thyroid carcinoma cells.

The present study aimed to investigate the effect of miR-449a-mediated Notch signaling pathway on the proliferation, apoptosis and invasion of papillary thyroid carcinoma cells. Human papillary thyroid carcinoma cell line TPC-1 was selected, and cells were grouped and transfected: Control group (without any treatment), negative control (NC) group (transfection with NC plasmid), miR-449a mimic group (transfection with miR-449a mimic), miR-449a inhibitor group (transfection with miR-449a inhibitor), DAPT group (addition of γ-secretase inhibitor DAPT to inhibit the Notch signaling pathway), and miR-449a inhibitor + DAPT group (transfection with miR-449a inhibitor and addition of DAPT). The target relationship between miR-449a and Notch1 was detected by dual-luciferase reporter assay. qRT-PCR and western blotting were used to assess the expression of miR-449a, Notch1 and Jagged1 in cells. Cell proliferation was detected using EdU; the cell cycle and apoptosis were detected by flow cytometry; cell invasion ability was detected by Transwell assay. PCNA, MMP-2, MMP-9, Bcl-2 and Bax mRNA and protein expression were assessed by qRT-PCR and western blotting. The results revealed that miR-449a negatively regulated Notch1. Compared with the control group, there was significantly increased miR-449a expression in the miR-449a mimic group, and there was significantly decreased expression of Notch1, Jagged1, PCNA, MMP-2, MMP-9 and Bcl-2, increased Bax, reduced cell proliferation, increased G1-phase cell fraction, decreased S-phase cell fraction, an increased apoptosis rate, and decreased invasion ability in the miR-449a mimic group and DAPT group (all P<0.05). However, the results in the miR-449a inhibitor group were the opposite of those in miR-449a mimic group (all P<0.05). There was no significant difference in cell proliferation, apoptosis and invasion in the NC group and miR-449a inhibitor + DAPT group compared to the control group (all P>0.05). miR-449a overexpression can inhibit Notch signaling pathway, thereby inhibiting the proliferation and invasion of papillary thyroid carcinoma cells and promoting cell apoptosis.

MicroRNA-92b suppresses proliferation of papillary thyroid cancer cells by regulating ilent information regulator 1 gene expression

Objective To investigate the regulating effects of microRNA (miRNA, miR)-122 on the expression of ilent information regulator 1 (SIRT1), and the proliferation abilities of papillary thyroid cancer cells. Methods Real-time quantitative polymerase chain reaction (Real-time PCR) was used to observe the expression of miR-122 in papillary thyroid carcinoma and adjacent tissues and to analyze its correlation with SIRT1 expression. After transfecting miR-122 mimics into human papillary thyroid carcinoma cell line TPC-1, the dual luciferase reporter gene assay was used to observe the regulation of miR-122 on SIRT1 gene. The effect of miR-122 on the proliferation of thyroid cancer cells was observed by CCK-8 assay. At the same time, the overexpressed SIRT1 plasmid was co-transfected into TPC-1 cells, and then the effect of miR-122 and SIRT1 expression on the proliferation of papillary thyroid carcinoma cells was detected by CCK-8 assay. Analysis was performed using GraphPad Prism 7.0 statistical software. Correlation analysis was performed using Spearman rank correlation analysis. Student’s t test was used to compare the two sample means. Results Compared with adjacent tissues (1.20±0.15), the expression level of miR-122 (3.06±0.26) in papillary thyroid carcinoma was significantly down-regulated, and the difference was statistically significant (t=5.972, P<0.01). Correlation analysis showed that miR-122 was negatively correlated with SIRT1 mRNA expression (r=-0.621, P<0.01). The relative luciferase activity of SIRT1 in miR-122 mimics transfection group was significantly lower than that in the control group, and the difference was statistically significant (t=17.730, P<0.01). The cell proliferation ability of miR-122 mimics group was significantly lower than that of the control group, and the difference was statistically significant (t=3.991, P<0.01). After co-transfection of the SIRT1 overexpression plasmid, the inhibition of the proliferation of thyroid papillary carcinoma cells by miR-122 mimics was reversed. Conclusion miR-122 can inhibit the proliferation of papillary thyroid cancer cells, and its mechanism may be related to the targeted inhibition of SIRT1 expression. Key words: MicroRNA-122; Ilent information regulator 1; Papillary thyroid cancer; Proliferation

Downregulation of microR-147b represses the proliferation and invasion of thyroid carcinoma cells by inhibiting Wnt/β-catenin signaling via targeting SOX15

microRNA-147b (miR-147b) is a newly identified tumor-related miRNA that is dysregulated in multiple cancer types. Yet, the role of miR-147b in thyroid carcinoma remains unknown. Herein, we found that miR-147b expression was upregulated in thyroid carcinoma tissues and cell lines. miR-147b inhibition decreased the proliferation, colony formation, and invasion of thyroid carcinoma cells. The tumor suppressive gene SRY-related high-mobility-group box gene 15 (SOX15) was predicted as a miR-147b target gene. SOX15 expression was markedly decreased in thyroid carcinoma tissues and inversely correlated with the miR-147b expression. SOX15 overexpression repressed the proliferation and invasion of thyroid carcinoma cells associated with downregulation of Wnt/β-catenin signaling. SOX15 knockdown abolished the miR-147b-inhibition-mediated antitumor effect. miR-147b inhibition or SOX15 overexpression retarded the tumor growth of thyroid carcinoma cells in vivo. Overall, our study suggests that miR-147b inhibition restrains the proliferation and invasion of thyroid carcinoma cells through upregulation of SOX15 and inhibition of Wnt/β-catenin signaling.

MiR-19a Is Involved In Progression And Malignancy Of Anaplastic Thyroid Cancer Cells.

BackgroundMicroRNAs (miRNAs) are endogenous, single-stranded, non-coding RNAs acting as negative regulators of gene expression involved in a number of physiological processes. MiRNAs’ expression is commonly dysregulated in many types of human tumor diseases and cancers, including thyroid cancers, and is often involved in tumor initiation and progression. miR-19a, a member of miR-17-92 cluster, has been demonstrated to promote cell growth in anaplastic thyroid cancer (ATC), the most advanced and aggressive thyroid cancer.PurposeIn this work, we investigate the potential contribution of miR-19a in thyroid cancer cells poor prognosis and de-differentiation.MethodsWe directly modulated the expression of miR-19a in papillary (PTC) and anaplastic thyroid carcinoma cell lines through transfection of specific miR-19a mimic or inhibitor. Further, we performed gene expression analysis of specific genes to evaluate miR-19a association with cell cycle, differentiation, and poor prognosis.ResultsOur data indicate that miR-19a overexpression in PTC cells significantly promotes cell growth, decreases the expression of differentiation genes and activates poor prognosis genes. Its inhibition in ATC cells reduces cell proliferation and the expression of genes related to poor prognosis but does not affect differentiation.ConclusionOur findings reveal the existence of functional associations between miR-19a expression and thyroid cancer progression and malignancy suggesting miR-19a as a novel candidate therapeutic target for ATC.

Uridine phosphorylase 1 associates to biological and clinical significance in thyroid carcinoma cell lines.

Thyroid cancer incidence has been continuity increasing worldwide. Uridine phosphorylase 1 (UPP1) is a protein‐coding gene and has been detected that UPP1 was the higher expression in many solid malignancies, just as head and neck cancers, breast cancer, compared with paired normal tissue. But the act of UPP1 in thyroid cancer is not explicit. In this article, we investigate the function of UPP1 expression in thyroid cancer. The Cancer Genome Atlas (TCGA) unpaired thyroid cancer and normal RNA‐seq data were downloaded, and our paired thyroid cancer and normal samples were analysed by a polymerase chain reaction. The expression of UPP1 was regulated by transfected small interfering RNA, and the function of UPP1 was determined via migration, invasion and cell proliferation assays. Western blot assay was achieved to determine the UPP1 expression correlates with the function of 5‐FU regulate epithelial‐mesenchymal transition. The significant upregulation of UPP1 in thyroid cancer tissues compared with normal thyroid tissues was revealed by our data and TCGA data. UPP1 overexpression was significantly correlated with lymph node metastasis, tumour stage and tumour size. In the cell, experiments showed that UPP1 low expression significantly suppressed the migration, invasion and proliferation. Western blot assay proves the effect of UPP1 expression on 5‐FU regulates epithelial‐mesenchymal transition pathway. UPP1 plays a crucial oncogene in thyroid cancer. Our findings indicate that UPP1 might be a biomarker of thyroid cancer and may act by regulating epithelial‐mesenchymal transition (EMT).

Long Non-coding RNA Expression in Anaplastic Thyroid Carcinomas.

Long non-coding RNAs (lncRNAs) participate in transcription and in epigenetic or post-transcriptional regulation of gene expression. They also have roles in epithelial to mesenchymal transition and in carcinogenesis. Because lncRNAs may also have a role in thyroid cancer progression, we examined a group of thyroid tumors which included papillary thyroid carcinomas and anaplastic thyroid carcinomas to determine the specific lncRNAs that were upregulated during thyroid tumor progression. An RT2 Profiler PCR Array Human Cancer Pathway Finder consisting of 84 lncRNAs (Qiagen) and fresh tissues of normal thyroid, PTCs, and ATCs with gene expression profiling was used to determine genes upregulated and downregulated in ATCs. Two of the most highly upregulated genes, prostate cancer antigen 3 (PCA3) and HOX antisense intergenic RNA myeloid 1 (HOTAIRM1 or HAM-1), were selected for further studies using a thyroid tissue microarray(TMA) with formalin-fixed paraffin-embedded tissues of normal thyroid (NT, n= 10), nodular goiters (NG, n= 10), follicular adenoma (FA, n= 32), follicular carcinoma (FCA, n= 28), papillary thyroid carcinoma (PTC, n= 28), follicular variant of papillary thyroid carcinoma (FVPTC, n = 28), and anaplastic thyroid carcinoma (ATC, n= 10). TMA sections were analyzed by in situ hybridization (ISH) using RNAscope technology. The results of ISH analyses were imaged with Vectra imaging technology and quantified with Nuance® and inForm® software. The TMA analysis was validated by qRT-PCR using FFPE tissues for RNA preparation. Cultured thyroid carcinoma cell lines (n= 7) were also used to analyze for lncRNAs by qRT-PCR. The results showed 11 lncRNAs upregulated and 7 downregulated lncRNAs more than twofold in the ATCS compared with PTCs. Two of the upregulated lncRNAs, PCA3 and HAM-1, were analyzed on a thyroid carcinoma TMA. There was increased expression of both lncRNAs in ATCs and PTCs compared with NT after TMA analysis. qRT-PCR analyses showed increased expression of both lncRNAs in ATCs compared with NT and PTCs. Analyses of these lncRNAs from cultured thyroid carcinoma cell lines by qRT-PCR showed the highest levels of lncRNA expression in ATCs. TGF-β treatment of cultured PTC and ATC cells for 21days led to increased expression of PCA3 lncRNA in both cell lines by day 14. These results show that the lncRNAs PCA3 and HAM-1 are upregulated during thyroid tumor development and progression and may function as oncogenes during tumor progression.

Interactions of Vascular Endothelial Growth Factor and p53 with miR-195 in Thyroid Carcinoma: Possible Therapeutic Targets in Aggressive Thyroid Cancers.

The clinical pathological features, as well as the cellular mechanisms of miR-195, have not been investigated in thyroid carcinoma. The aim of this study is to identify the interactions of vascular endothelial growth factor (VEGF), p53 and miR-195 in thyroid carcinoma. The clinical and pathological features of miR-195 were also investigated. The expression levels of miR-195 were identified in 123 primary thyroid carcinomas, 40 lymph nodes with metastatic papillary thyroid carcinomas and seven non-neoplastic thyroid tissues (controls) as well as two thyroid carcinoma cell lines, B-CPAP (from metastasizing human papillary thyroid carcinoma) and MB-1 (from anaplastic thyroid carcinoma), by the real-time polymerase chain reaction. Using Western blot and immunofluorescence, the effects of exogenous miR-195 on VEGF-A and p53 protein expression levels were examined. Then, cell cycle and apoptosis assays were performed to evaluate the roles of miR-195 in cell cycle progression and apoptosis. The expression of miR-195 was downregulated in majority of the papillary thyroid carcinoma tissue as well as in cells. Introduction of exogenous miR-195 resulted in downregulation of VEGF-A and upregulation of p53 protein expressions. Upregulation of miR-195 in thyroid carcinoma cells resulted in cell cycle arrest. Moreover, we demonstrated that miR-195 inhibits cell cycle progression by induction of apoptosis in the thyroid carcinoma cells. Our findings showed for the first time that miR-195 acts as a tumour suppressor and regulates cell cycle progression and apoptosis by targeting VEGF-A and p53 in thyroid carcinoma. The current study exhibited that miR-195 might represent a potential therapeutic target for patients with thyroid carcinomas having aggressive clinical behaviour.