Papillary Thyroid Cancer Cell Proliferation Research Articles

IL-34 is a potential biomarker for the treatment of papillary thyroid cancer.

BackgroundInterleukin (IL)‐34 is a recently discovered pro‐inflammatory cytokine and is a vital regulator in different tumor types. However, the function of IL‐34 in thyroid carcinoma has yet to be investigated. In this study, we analyzed the expression of IL‐34 in human papillary thyroid cancer (PTC) samples and determined its effects on the proliferation and apoptosis of PTC cells.MethodsWe examined the expression of IL‐34 in serum and tissue samples of patients with PTC by Western blotting and ELISA assay and analyzed its association with clinicopathological features including tumor size, tumor node metastasis (TNM) stage, and lymph node metastasis (LNM). We selected TPC1 and K1 for knockdown or overexpressing of IL‐34 via small interference RNA transfection. The proliferation of PTC cells was evaluated by CCK8 assay. We further investigated the role of IL‐34 in apoptosis by flow cytometry and studied the protein levels of epithelial‐mesenchymal transition (EMT) biomarkers, phosphorylated extracellular‐regulated kinase (ERK), and total‐ERK (t‐ERK) by Western blotting.ResultsOur results show that IL‐34 is significantly upregulated in serum and tissue samples from patients with PTC. IL‐34 promotes the proliferation and suppresses apoptosis in PTC cells. In addition, IL‐34 can promote the EMT and activate ERK signaling pathway in PTC cells.ConclusionThis study provides novel evidence that IL‐34 serves as an oncogene in PTC. IL‐34 promotes proliferation, EMT phenotype, and ERK signaling pathway and inhibits apoptosis in PTC cells. Therefore, IL‐34 may be a potent therapeutic target for the treatment of PTC.

Linc00324 promotes the progression of papillary thyroid cancer via regulating Notch signaling pathway.

To explore the expression of linc00324 in papillary thyroid cancer (PTC) and its effect on the biological function of PTC cells. A total of 60 pairs of PTC and para-carcinoma normal tissues surgically excised were collected. The expression of linc00324 in PTC tissues and cells was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR), and the expression of linc00324 in PTC cells was silenced using the small-interfering RNA (siRNA). Then, the effects of linc00324 on the PTC cell proliferation, apoptosis, and cycle and the downstream Notch signaling pathway were determined via methyl thiazolyl tetrazolium (MTT) assay, colony formation assay, flow cytometry, and Western blotting, respectively. The expression of linc00324 was upregulated in 48 out of 60 cases of PTC tissues, and it was increased in PTC cells compared with that in human thyroid follicular epithelial cells Nthy-ori 3-1. The results of MTT assay and colony formation assay showed that the proliferation of PTC cells declined after interference in linc00324 expression. The findings of flow cytometry revealed that the cell cycle was arrested in G1/G0 phase with a higher apoptosis rate in si-linc00324 group compared with that in the si-NC group. According to the data of Western blotting, the molecular markers for the downstream Notch signaling pathway were altered after interference in linc00324 expression. The expression of linc00324 is significantly increased in PTC tissues and cells. Silencing linc00324 may inhibit the proliferation of PTC cells, arrest the cell cycle in G1/G0 phase, and promote the apoptosis by inhibiting the Notch signaling pathway.

MON-536 SPOCK1 Promotes the Progression of Papillary Thyroid Cancer ViaPI3K/Akt Signaling Activation

With the increasing incidence, thyroid cancer as one of the most common malignancy, has got widespread attention during the past few years. Papillary thyroid cancer (PTC) is the most common thyroid cancer type. Understanding the underlining molecular mechanisms of PTC is of great interest. The oncogenic role of SPARC/osteonectin, cwcv, and kazal-like domain proteoglycan 1 (SPOCK1) has been demonstrated in several cancers, however, the clinical and functional significance of SPOCK1 in PTC are largely unknown. Here, we found that the expression of SPOCK1 was upregulated in PTC tissues when comparing with the adjacent normal thyroid tissues. The overexpression of SPOCK1 was associated with the clinicopathological characteristics of the patients with PTC. We demonstrated that the proliferation of PTC cells was significantly promoted and the apoptosis of PTC cells was significantly inhibited in cells with overexpression of SPOCK1. Furthermore, we showed that knockdown of SPOCK1 arrested the cell cycle in G0/G1 phase and promoted the apoptosis in PTC cell lines. Importantly, our data suggested that SPOCK1 promoted the progression of PTC cell via regulating the PI3K/Akt signaling pathway. Taking together, our findings demonstrate that SPOCK1 enhances the activation of PI3K/Akt signaling pathway, thereby promoting the proliferation and inhibiting the apoptosis of PTC cells.

MiR-200c promotes proliferation of papillary thyroid cancer cells via Wnt/β-catenin signaling pathway.

To investigate the potential effects of miR-200c on proliferation and apoptosis of papillary thyroid cancer (PTC) cells. Micro ribonucleic acid-200c (miR-200c) inhibitor was transfected to down-regulate miR-200c expression. Cell counting kit-8 (CCK-8), colony formation experiment, and flow cytometry were used to detect the effects of miR-200c knockdown on proliferation and apoptosis of Butylated Hydroxytoluene 101 (BHT101) cells. The dual-luciferase reporter gene assay was conducted to detect whether miR-200c directly binds to the target gene. After knocking down miR-200c, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting analysis were performed to detect changes of target genes regarding messenger RNA (mRNA) and protein. Western blotting analysis was also adopted to detect gene expression of Wnt/β-catenin signaling pathway-related proteins. Compared with those in control group, the proliferation and clone formation ability of BHT101 cells in miR-200c knockdown group were significantly inhibited (p<0.05), while the apoptosis rate increased markedly (p<0.05). Dachshund Family Transcription Factor 1 (DACH1) was the direct target gene of miR-200c. After miR-200c knockdown, the expression levels of Wnt/β-catenin signaling pathway members (including c-Myc, β catenin and cyclin D1) all decreased. MiR-200c is a tumor suppressor miRNA, which promotes proliferation of PTC cells and activates Wnt/β-catenin signaling pathway by directly regulating the corresponding target protein, DACH1.

High expression of LINC0163 promotes progression of papillary thyroid cancer by regulating epithelial-mesenchymal transition MITF

The purpose of this study was to detect the expression of long non-coding ribonucleic acid 00163 (LINC00163) in human papillary thyroid cancer (PTC), and to observe the influence of downregulated LINC00163 on the proliferative and metastatic capacities of human PTC cells. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) assay was applied to measure the expression level of LINC00163 in PTC tissues and para-carcinoma tissues, as well as that in normal human thyroid cells (Nthy-ori3-1) and PTC cells. After the expression of LINC00163 in PTC cells was interfered, qRT-PCR assay was performed to determine the interference efficiency, and colony formation and Cell Counting Kit-8 (CCK-8) assays were conducted to study the impacts of small interfering (si)-LINC00163 on the proliferative capacity of PTC cells. Moreover, wound healing and transwell assays were adopted to investigate the changes in the migratory and invasive abilities of PTC cells after the interference in the expression of LINC00163 in PTC cells. Finally, the changes in expressions of molecular markers in downstream signaling pathways after interference in LINC00163 expression were examined via Western blotting assay. In 51 cases of PTC tissues and corresponding para-carcinoma tissues, 41 cases exhibited an up-regulated expression of LINC00163, and qRT-PCR results indicated that PTC cells also had an up-regulated expression of LINC00163 compared with normal human thyroid cells. After the expression of LINC00163 in PTC cells was interfered, the results of colony formation and CCK-8 assays manifested that the proliferative capacity of the cells declined. It was also shown in wound-healing and transwell assay results that the migratory and invasive abilities of the cells were weakened. In addition, the results of Western blotting assay revealed expression changes in the molecular markers of epithelial-mesenchymal transition (EMT). The expression of LINC00163 in NSCLC tissues and cells is upregulated, and highly expressed LINC00163 can promote PTC cell proliferation and metastasis by regulating the EMT.

MiR-146b-5p Regulates the Expression of Long Noncoding RNA MALAT1 and Its Effect on the Invasion and Proliferation of Papillary Thyroid Cancer.

Background: The incidence of thyroid cancer has increased dramatically in recent decades due, in large part, to identifications of subclinical diseases. Literature on thyroid cancer has examined the pathogenesis of high invasive papillary thyroid cancer (PTC) and has improved the prevention and treatment of PTC. This study aims to investigate the effects of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) on PTC migration and invasion, and clarify the regulatory mechanisms between miR-146b-5p and MALAT1. Materials and Methods: In this study, we examined the differential expression of MALAT1, miR-146b-5p, and DNA methyltransferases 3A (DNMT3A) in PTC tissues. The effect of MALAT1 on the proliferation and invasion ability of PTC cells was verified by constructing a sh-MALAT1 knockdown cell model. Correlations between MALAT1, miR-146b-5p, and DNMT3A were analyzed by the Pearson correlation method. Finally, we verified the regulatory relationship between miR-146b-5p and MALAT1 by the luciferase assay and rescue assay. Results: The expression of MALAT1 was upregulated in PTC tissues and cells, while a MALAT1 knockdown counteracted cellular activity, migration, and invasion of B-CPAP and K1 cells. The relationship between miR-146b-5p and DNMT3A was negative, while the relationship between miR-146b-5p and MALAT1 was positive. Both genes were separately detected using the Pearson correlation method. The luciferase assay and rescue assay demonstrated that a binding site in miR-146b-5p was existent in the 3' untranslated region of DNMT3A, while a knockdown of DNMT3A partially rescued si-miR-146b-5p induced proliferation, migration, and invasion effects on PTC cells. Conclusions: The MALAT1 gene is highly expressed in PTC, while the knockdown MALAT1 gene attenuates the cellular activity and invasive ability of PTC cells. The microRNA miR-146b-5p can promote a MALAT1 expression by negatively regulating DNMT3A in PTC.

BAG5 promotes invasion of papillary thyroid cancer cells via upregulation of fibronectin 1 at the translational level

Papillary thyroid cancer (PTC), the most common thyroid malignancy, has a strong propensity for neck lymph node metastasis, which will increase the risk of local recurrence and decrease the survival in some high-risk groups. Hence, it is essential to set up a reliable biomarker to predict lymph node metastasis. BAG5 is a unique member of the BAG cochaperone family because it consists of more than one BAG domain, which acts as modulator of chaperone activity. In this study, we found that expression of BAG5 was significantly increased in PTC cells and tissues. Neither overexpression nor downregulation of BAG5 altered the proliferation of PTC cells. On the contrary, overexpression of BAG5 significantly promoted, while knockdown of BAG5 significantly decreased migration and invasion of PTC cells. Along with this, fibronectin 1 (FN1) was significantly increased and decreased in cells that overexpress or downregulate BAG5, respectively. Mechanistically, we found that BAG5 modulated FN1 expression at the translational level and promoted invasion via suppression of miR-144-3p, which targeted the 3′ untranslational region (UTR) of FN1 transcript. This study suggests that BAG5 is an important regulator of migration and invasion in PTC cells and may represent a novel therapeutic target for intervening in PTC progression.

RETRACTED ARTICLE: The HOTAIRM1/miR-107/TDG axis regulates papillary thyroid cancer cell proliferation and invasion

The long noncoding RNA (lncRNA), HOX antisense intergenic RNA myeloid 1 (HOTAIRM1), has been shown to act as a tumor suppressor in various human cancers. However, the overall biological roles and clinical significance of HOTAIRM1 in papillary thyroid cancer (PTC) have not been investigated. In this study, we used quantitative reverse transcription PCR (qRT-PCR) to show that HOTAIRM1 was significantly downregulated in PTC tissues and low HOTAIRM1 expression levels were associated with lymph node metastasis and advanced TNM stage. We performed Cell Counting Kit-8, plate colony-formation, flow cytometric apoptosis, transwell, and scratch wound healing assays. Overexpression of HOTAIRM1 was found to inhibit PTC cell proliferation, invasion, and migration in vitro. Additionally, we identified miR-107 as a target of HOTAIRM1 using online bioinformatics tools. Dual-luciferase reporter gene and RNA immunoprecipitation assays were used to confirm that HOTAIRM1 acted as a competing endogenous RNA of miR-107. Furthermore, enhancement of miR-107 could potentially reverse the effects of HOTAIRM1 overexpression in vitro. Inhibition of miR-107 suppressed PTC cell proliferation, invasion, and migration in vitro. HOTAIRM1 overexpression and miR-107 inhibition impaired tumorigenesis in vivo in mouse xenografts. Bioinformatics prediction and a dual-luciferase reporter gene assay demonstrated the binding between miR-107 and the 3′-untranslated region of TDG. The results of qRT-PCR and western blotting assays suggested that HOTAIRM1 could regulate the expression of TDG in an miR-107-meditated manner. In conclusion, we validated HOTAIRM1 as a novel tumor-suppressor lncRNA in PTC and proposed that the HOTAIRM1/miR-107/TDG axis may serve as a therapeutic target for PTC.

Sanguinarine Induces Apoptosis in Papillary Thyroid Cancer Cells via Generation of Reactive Oxygen Species.

Sanguinarine (SNG), a natural compound with an array of pharmacological activities, has promising therapeutic potential against a number of pathological conditions, including malignancies. In the present study, we have investigated the antiproliferative potential of SNG against two well-characterized papillary thyroid cancer (PTC) cell lines, BCPAP and TPC-1. SNG significantly inhibited cell proliferation of PTC cells in a dose and time-dependent manner. Western blot analysis revealed that SNG markedly attenuated deregulated expression of p-STAT3, without affecting total STAT3, and inhibited growth of PTC via activation of apoptotic and autophagy signaling cascade, as SNG treatment of PTC cells led to the activation of caspase-3 and caspase-8; cleavage of PARP and activation of autophagy markers. Further, SNG-mediated anticancer effects in PTC cells involved the generation of reactive oxygen species (ROS) as N-acetyl cysteine (NAC), an inhibitor of ROS, prevented SNG-mediated antiproliferative, apoptosis and autophagy inducing action. Interestingly, SNG also sensitized PTC cells to chemotherapeutic drug cisplatin, which was inhibited by NAC. Finally, SNG suppressed the growth of PTC thyrospheres and downregulated stemness markers ALDH2 and SOX2. Altogether, the findings of the current study suggest that SNG has anticancer potential against PTC cells as well its derived cancer stem-like cells, most likely via inactivation of STAT3 and its associated signaling molecules.

MiR-1256 inhibits cell proliferation and cell cycle progression in papillary thyroid cancer by targeting 5-hydroxy tryptamine receptor 3A.

Aberrant expression of miR-1256 has been reported to be closely associated with the development and progression of tumors, including colon cancer and lung cancer. However, study of its expression pattern and functional role in papillary thyroid cancer (PTC) is rare. Using quantitative real time PCR analysis, we found miR-1256 was significantly down-regulated in PTC tissues and cell lines. The correlation of miR-1256 expression with clinicopathological features was statistically analyzed. The results showed miR-1256 expression was significantly correlated with tumor size (p = 0.0124) and TNM stage (p = 0.0032). Restoring miR-1256 expression significantly inhibited proliferation and cell cycle progression of PTC cells demonstrated by CCK-8 and flow cytometry assays. Luciferase reporter assay and biotin-avidin pull-down assay showed miR-1256 can directly target 5-hydroxytryptamine receptor 3A (HTR3A) in PTC cells. The expression of miR-1256 was inversely correlated with HTR3A expression in PTC tissues. Knockdown of HTR3A imitated the suppressive effects of miR-1256 in PTC cells. Ectopic expression of HTR3A can antagonize the effects of miR-1256 on PTC cells. Furthermore, the suppressive effects of miR-1256 on the expression of PCNA, CDK4, Cyclin D1, and p21 were partially reversed by HTR3A overexpression in PTC cells. In summary, our data suggested that miR-1256 could suppress PTC cellular function by targeting HTR3A, which might be a potential therapeutic target for patients with PTC.

S100A12 is a promising biomarker in papillary thyroid cancer

S100A12 belongs to the S100 family and acts as a vital regulator in different types of tumors. However, the function of S100A12 in thyroid carcinoma has not yet been investigated. In this study, we analyzed the expression of S100A12 in human papillary thyroid cancer (PTC) samples and two PTC cell lines. In addition, we explored the effects of S100A12 on PTC cell progression in vitro and in vivo. Our results showed that S100A12 was significantly upregulated in PTC specimens. Moreover, silencing S100A12 markedly inhibited PTC cell proliferation, migration, invasion and cell cycle progression. In addition, knockdown of S100A12 significantly reduced the expression of CyclinD1, CDK4 and p-ERK in PTC cells. An in vivo study also showed that silencing S100A12 dramatically suppressed tumor cell growth and decreased Ki67 expression in a xenograft mouse model. This study provides novel evidence that S100A12 serves as an oncogene in PTC. Knockdown of S100A12 suppressed PTC cell proliferation, migration, and invasion and induced G0/G1 phase arrest via the inhibition of the ERK signaling pathway. Therefore, S100A12 may be a potent therapeutic target for PTC.

&lt;p&gt;miR-873-5p Inhibits Cell Migration and Invasion of Papillary Thyroid Cancer via Regulation of CXCL16&lt;/p&gt;

AimPapillary thyroid cancer (PTC) is the most common type of thyroid cancer with an increasing morbidity. MicroRNAs (miRNAs) play the pivotal roles in PTC occurrence and development. The aim of this study was to investigate the biological functions of miR-873-5p and its underlying molecular mechanisms in PTC.MethodsReverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis was performed to detect miR-873-5p expressions in PTC tissues and cell lines. The target gene of miR-873-5p was predicted by TargetScan and confirmed by dual-luciferase reporter assay. Furthermore, cell proliferation, migration and invasion were assessed by CCK-8, wound healing assay and transwell assay, respectively. Additionally, the expressions of CXCL16, MMP1, MMP9 and MMP13 were measured by RT-qPCR and Western blot methods, and p65, Rel-B and their phosphorylation levels were examined by Western blot.ResultsWe found that miR-873-5p expression was downregulated in PTC tissues and cell lines. Moreover, CXCL16 was identified as a target of miR-873-5p, and its expression was upregulated in PTC tissues and cells at both mRNA and protein levels. Functionally, overexpression of miR-873-5p inhibited PTC cell proliferation, migration and invasion, while co-transfection of CXCL16 overexpression plasmid reversed the anti-tumor behaviors induced by miR-873-5p. In addition, miR-873-5p overexpression suppressed the phosphorylation of p65 and Rel-B, and decreased the mRNA and protein expression of MMP1, MMP9 and MMP13, while overexpression of CXCL16 partially abrogated the effects of miR-873-5p.ConclusionMiR-873-5p functions as a tumor suppressor in PTC by inhibiting the proliferation, migration and invasion of the PTC cells via targeting CXCL16. These findings might provide a potential novel target for the therapy of PTC.

Long noncoding RNA CASC9 promotes the proliferation and metastasis of papillary thyroid cancer via sponging miR-488-3p.

Cancer susceptibility candidate 9 (CASC9) is a recently identified lncRNA that acted as a tumor promotor in diversified cancer types. However, its role in papillary thyroid cancer (PTC) remains unknown. The expression of CASC9 was measured in 52 human PTC tissues and PTC cell lines as well as their controls. The proliferation, migration, and invasion of PTC cells were determined after knockdown or overexpression of CASC9 to evaluate the effect of CASC9 on PTC cells. Also, the role of PTC tumorigenesis was confirmed in mice xenograft models. Additionally, the underlying mechanisms of CASC9 were further researched. We found that CASC9 expression was augmented in human PTC tissues and cells. Higher CASC9 expression was associated with large tumor size, advanced stage, or lymph node metastasis. Downregulation of CASC9 significantly attenuated the proliferative, migrative, and invasive abilities of PTC cells, and suppressed tumorigenesis in vivo. While overexpression of CASC9 elevated the proliferation, migration, and invasion of PTC cells. miR‐488‐3p expression was decreased, and ADAM9 level was increased in PTC tissues and cells. CASC9 expression was negatively related to miR‐488‐3p, but positively associated with ADAM9 expression in PTC tissues. Molecular mechanism analysis revealed that CASC9 functioned via sponging miR‐488‐3p to regulate ADAM9 expression, followed by activation of EGFR‐Akt signaling. In conclusion, lncRNA CASC9 promoted the malignant phenotypes of PTC via modulating miR‐488‐3p/ADAM9 pathway. This study may provide a novel therapeutic target for the treatment of PTC.

TRIM30 modulates Interleukin-22-regulated papillary thyroid Cancer cell migration and invasion by targeting Sox17 for K48-linked Polyubiquitination

BackgroundInterleukin-22 (IL-22) belongs to the IL-10 cytokine family and is mainly produced by activated Th1 cells. Although IL-22 expression is reported to be elevated in many cancers, and increased IL-22 expression correlates with tumor progression and poor prognosis, little is known about the role of IL-22 in papillary thyroid cancer (PTC). We previously demonstrated that IL-22 promotes PTC cell migration and invasion through the microRNA-595/Sox17 axis.MethodsWe used qRT-PCR and western blot to determine TRIM30, Sox17 and β-catenin expression in PTC cells. Knockdown and overexpression were performed to detect the role of TRIM30/Sox17/β-catenin axis on the migration and invasion PTC cells. Co-IP were used to determine the interaction between TRIM30 and Sox17.FindingsIn this study, we demonstrated that IL-22 triggered tripartite-motif protein 30 (TRIM30) association with Sox17, thereby mediating K48-linked polyubiquitination of Sox17. We then demonstrated that TRIM30 was a positive regulator of IL-22-regulated migration and invasion of PTC cells. We also found that IL-22 induced the transcriptional activity of β-catenin and translocation of β-catenin from cytosol to the nucleus. Upon investigating the mechanisms behind this event, we found that IL-22 disrupted Sox17/β-catenin interactions by inducing TRIM30/Sox17 interactions, leading to promotion of β-catenin-dependent signaling. The analysis of hundreds of clinical specimens revealed that IL-22, TRIM30 and β-catenin levels were upregulated in PTC tissues compared with normal thyroid, and that their expression levels were closely correlated. Taken together, under the influence of IL-22, by sequestration of Sox17, TRIM30 promotes β-catenin-dependent signaling that promotes PTC cell proliferation.

MicroRNA-675 directly targets MAPK1 to suppress the oncogenicity of papillary thyroid cancer and is sponged by long non-coding RNA RMRP.

BackgroundMicroRNA-675-5p (miR-675-5p) is dysregulated in multiple human cancers, but its involvement in papillary thyroid cancer (PTC) remains to be investigated. This study aimed to examine the expression pattern of miR-675 in PTC, determine the effects of miR-675 on regulating the progression of PTC, and to explore the underlying molecular mechanisms.MethodsThe expression profile of miR-675 in PTC tissues and cell lines was determined using RT-qPCR. CCK-8, transwell migration and invasion assays, and xenograft tumors in nude mice were employed to analyze proliferation, in vitro migration and invasion, and in vivo tumor growth of PTC cells, respectively. The putative target of miR-675 was predicted using bioinformatic algorithms and was confirmed using luciferase reporter assays, RT-qPCR, and Western blotting.ResultsmiR-675 expression was decreased in PTC tissues and cell lines. A low level of miR-675 expression was significantly correlated with lymphatic metastasis and TNM stage in PTC patients. Ectopic miR-675 expression suppressed PTC cell proliferation, migration, and invasion in vitro and hindered tumor growth in vivo. Mitogen-activated protein kinase 1 (MAPK1) was found to be the direct target gene of miR-675 in PTC cells. MAPK1 reintroduction negated the tumor-suppressing effect of miR-675 overexpression in PTC cells. Furthermore, the lncRNA mitochondrial RNA processing endoribonuclease (RMRP) functioned as a ceRNA of miR-675 in PTC cells. Silencing RMRP expression inhibited the growth and metastasis of PTC cells by sponging miR-675 and regulating MAPK1.ConclusionThese findings revealed that miR-675 directly targets MAPK1 and is sponged by lncRNA RMRP to inhibit the oncogenicity of PTC, suggesting the RMRP-miR-675-MAPK1 pathway is an effective target for the treatment of PTC patients.

Hsa_circ_0058124 promotes papillary thyroid cancer tumorigenesis and invasiveness through the NOTCH3/GATAD2A axis

BackgroundDespite a good and overall prognosis, papillary thyroid cancer (PTC) can still affect the quality of life of many patients, and can even be life-threatening due to its invasiveness and metastasis. Emerging studies demonstrate that circular RNAs (circRNAs) participate in the regulation of various cancers. However, the circRNA profile in invasive PTC is still not well understood.MethodsCompeting endogenous RNA (ceRNA) microarrays were performed to determine circRNAs contributed to the tumorigenesis and invasiveness of PTC. Bioinformatics methods were used to narrow down the candidate circRNAs. Quantitative real-time polymerase chain reaction (qRT-PCR) assays revealed a significant upregulation of hsa_circ_0058124 in PTC tissue and a close correlation with a poor prognosis for PTC patients. RNA fluorescence in situ hybridization and Cell fractionation assay were used to investigate the subcellular location of hsa_circ_0058124. Then, we examined the functions of hsa_circ_0058124 in PTC by cell proliferation, cell cycle, apoptosis, migration and invasion assay. Mechanistically, RNA sequencing and GSEA analysis were applied to predict the downstream pathway of hsa_circ_0058124. Dual-luciferase report assays were used to explore the potential miRNA sponge role of hsa_circ_0058124. Western blotting, cell proliferation, cell cycle, cell apoptosis, migration and invasion, and mouse xenograft assay were used to validate the effects of hsa_circ_0058124/NOTCH3/GATAD2A axis on PTC progression.ResultsIn the current study, a novel hsa_circ_0058124 on 2q35 was identified and explored in PTC. Hsa_circ_0058124 is associated with the malignant features and poor outcomes of PTC patients. Hsa_circ_0058124 acts as an oncogenic driver that promotes PTC cell proliferation, tumorigenicity, tumor invasion, and metastasis, which functions as a competing endogenous RNA to modulate miRNA-218-5p and its target gene NUMB expression, and consequently with repression of the NOTCH3/GATAD2A signaling axis in vitro and in vivo.ConclusionsThis study unveils a novel biomarker panel consisting of the hsa_circ_0058124/NOTCH3/GATAD2A axis which is critical for PTC tumorigenesis and invasiveness and may represent a novel therapeutic target for intervening in PTC progression.

MiR-4500 Regulates PLXNC1 and Inhibits Papillary Thyroid Cancer Progression.

Although most patients with papillary thyroid cancer (PTC) are curable, there are still a few patients showing poor outcomes and increased risk of secondary cancers after therapies. In this study, we aimed to investigate the correlation between miR-4500 and PTC and to explore its molecular functions. A total of 50 patients were included, and sonography and histological examinations were used for diagnosis. Quantitative real-time polymerase chain reaction (qRT-PCR) was applied for detection of mRNA levels while Western blotting was used for measuring protein expression. Cell proliferation was tested using CCK-8 and colony formation assays. Caspase-3 activity and nucleosomal fragmentation assays were employed to test cell apoptosis. Cell invasive ability was measured using transwell assay. MiR-4500 target was identified using luciferase assay and RNA pull-down assay. MiR-4500 expression was significantly decreased in five PTC cell lines compared with Nthy-ori 3-1 cells and in PTC tissues compared with adjacent normal thyroid tissues, respectively. Decreased expression of miR-4500 showed lower survival rate, higher cancer stage, and lymphatic metastasis. Therefore, our results implied that miR-4500 could serve as a potential biomarker for PTC prognosis. Overexpression of miR-4500 repressed colony formation, proliferation, and invasiveness of PTC cells whereas increased cell apoptosis. We identified that PLXNC1 was a direct target of miR-4500. PLXNC1 knockdown showed similar effects on cell viability, colony formation, and cell apoptosis as overexpression of miR-4500 in PTC cells. In conclusion, miR-4500 inhibits the malignant transformation of PTC cells by directly targeting and repressing PLXNC1.

FOXE1 inhibits cell proliferation, migration and invasion of papillary thyroid cancer by regulating PDGFA

PurposeForkhead box E1 (FOXE1) plays an important role in the development, proliferation and differentiation of thyroid cells. However, the biological functions of FOXE1 in papillary thyroid cancer (PTC) remain unclear. Materials and methodsIn this study, the level of FOXE1 expression was examined in human PTC tissues and cells. Then, the high expression of FOXE1 was specifically silenced by RNA interference in vitro. Subsequently, FOXE1-shRNA was transfected into PTC cells (TPC-1 and K1). The effects on cell proliferation, migration and invasion were evaluated. In addition, FOXE1 targets were screened by cDNA microarray assays. The correlation between the expression of target gene platelet-derived growth factor A (PDGFA) and clinicopathological features of PTC patients was analysed. ResultsFOXE1 is highly expressed in PTC tissues and PTC cell lines. The silencing of FOXE1 significantly promotes PTC cell proliferation, migration and invasion in vitro. The cDNA microarray analyses show that PDGFA is a critical downstream target gene of FOXE1 in PTC cells. It was also observed that PDGFA is negatively regulated by FOXE1 in PTC. The clinical data indicate that the low expression level of PDGFA is correlated with the small size of PTC. ConclusionCollectively, the results indicate for the first time that high expression of FOXE1 may function as a tumour suppressor in the early stage of PTC and restrain the proliferation, migration and invasion of PTC by negatively regulating PDGFA expression. Thus, FOXE1 could serve as a prognostic biomarker for PTC.

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.