Induction Toripalimab and Chemotherapy for Organ Preservation in Locally Advanced Laryngeal and Hypopharyngeal Cancer: A Single-Arm Phase II Clinical Trial.

PURPOSE: The aim of this study was to assess the efficacy, toxicities, and potential role of larynx preservation of induction chemotherapy combined with programmed cell death protein 1 (PD-1) inhibitor in locally advanced laryngeal and hypopharyngeal cancer. PATIENTS AND METHODS: This is a single-arm phase II study. Patients with histopathologically confirmed, resectable locally advanced laryngeal/hypopharyngeal squamous cell carcinoma and Eastern Cooperative Oncology Group Performance Status 0-1 were eligible. Three cycles of induction chemotherapy (paclitaxel 175 mg/m2 d1, cisplatin 25 mg/m2 d1-3) combined with PD-1 inhibitor (toripalimab 240 mg d0) were administered. Response assessment was performed after induction chemoimmunotherapy using RECIST 1.1 criteria. Patients with a complete/partial response of the primary tumor received concurrent chemoradiation, followed by maintenance therapy of toripalimab. Otherwise, patients were referred to surgery, followed by adjuvant (chemo) radiation and maintenance therapy of toripalimab. The primary endpoint is a larynx preservation rate at 3 months postradiation. RESULTS: Twenty-seven patients were enrolled. Most cases exhibited stage IV disease (81.5%), with T4 representing 37.0%. Five patients underwent pretreatment tracheostomy because of impaired larynx function. Overall response rate of induction chemoimmunotherapy was 85.2%. At 3 months postradiation, the larynx preservation rate was 88.9%. With a median follow-up of 18.7 months, the 1-year overall survival rate, progression-free survival rate, and larynx preservation rate were 84.7%, 77.6%, and 88.7%, respectively. When excluding those with pretreatment tracheostomy, the 1-year larynx preservation rate was 95.5%. Exploratory analysis revealed that relapse correlated with enrichment of RNA signature of hypoxia and M2 macrophage-associated genes. CONCLUSIONS: Induction toripalimab combined with chemotherapy provided encouraging activity, promising larynx preservation rate and acceptable toxicity in this cohort of extensively locally advanced laryngeal and hypopharyngeal cancer.

Prognostic evaluation model for papillary thyroid cancer: a retrospective study of 660 cases.

BACKGROUND: Prognostic evaluation model for papillary thyroid cancer is very important for guiding the personalized treatment and follow-up strategy. There are imperfections in the system existed, and there is no suitable prognostic model for Chinese population. METHODS: This study was based on the clinic and follow-up data of 660 patients received surgical treatments in the Department of Head and Neck Surgery, Fudan University Shanghai Cancer Center from 2000 to 2005. Cox univariate/multivariate analysis was used to explore the influence factors of prognosis, and nomogram model was performed to establish a prognostic prediction system. RESULTS: Totally, 660 patients for initial treatment were included in our analysis with a median follow-up of 113.5 months. Five-, 10- and 15-year disease-free survival rate was 95.5%, 90.2% and 89.2%. Five-, 10- and 15-year overall survival rate was 99.7%, 99.2% and 99.1%. Residual tumor was associated with overall survival [hazard ratio (HR) 20.9, 95% confidence interval (CI): 2.3-187.6, P<0.05]. Age of onset (HR 2.00, 95% CI: 1.17-3.42, P<0.05) and the dimension of lymph nodes involved (0.2-3 cm: HR 3.67, 95% CI: 1.13-11.87, P<0.05; >3 cm: HR 5.20, 95% CI: 1.31-20.65, P<0.05) were independent influence factors of disease-free survival. The nomogram model for predicting prognosis of papillary thyroid cancer was established with a moderate predictive value (c-index 0.71, 95% CI: 0.57-0.84). CONCLUSIONS: The prognosis of papillary thyroid cancer is very good after appropriate treatment. Age and the dimension of lymph nodes involved were independent influence factors of disease-free survival for papillary thyroid cancer. A prognostic prediction model for Chinese population was established with moderate predictive value. A study with larger samples and including more factors of prognosis is necessary to increase the predictive value of model.

Multiple lectin assays in detecting glycol-alteration status of serum NRG1 in papillary thyroid cancer.

BACKGROUND: Neuregulin 1 (NRG1) is a membrane glycoprotein mediating cell-to-cell signaling and has a crucial role in the growth and development of various organ systems. Our study explored its diagnostic value in distinguishing BRAF V600E mutant status in papillary thyroid cancer (PTC) patients by analyzing multiple glycan patterns of serum NRG1 through lectin assays. METHODS: We first extracted serum from PTC patients and tested BRAF V600E mutation by immunohistochemical (IHC) staining. Then we applied antibody overlay lectin microarray and lectin blot to detect glycol-alterations of NRG1. Then Aleuria aurantia lectin (AAL) ELISA was performed according to ELISA index to test the protein fucosylation level of NRG1 (Fuc-NRG1). RESULTS: We got glycan profiles of 14 lectins, including GNL, GSL2, AAL, BPL, ECL, CAL, NML, HHL, PHA-L, RCA-I, ConA, DBA, PWA and LEL. Six of them, namely, GSL2, BPL, NML, HHL, PHA-L and LEL, had significantly increased binding affinity capacity in BRAF(+) PTC compared with BRAF(-) PTC controls. LEL, BPL and NML tended to bind to NRG1 in BRAF(+) PTC group. Both AAL ELISA and protein ELISA assays showed that the fucosylated structures of NRG1 had a remarkable increase in BRAF V600E mutant PTC patients compared with BRAF wild type PTC controls. CONCLUSIONS: This study sheds a new light on the role of NRG1 glycosylation in PTC. NRG1 could serve as a supplementary glycobiomarker for BRAF indicator in discrimination of PTC patients with BRAF wild type negative fine needle aspiration results.

Prognostic and clinical significance of HOXC9 and HOXD10 in papillary thyroid cancer.

BACKGROUND: The homebox superfamily play an important role in tumorigenesis. HOXC9 and HOXD10 were reported playing critical roles in tumor progression in many malignant tumors. This study aimed to research the expression of HOXC9 and HOXD10 in papillary thyroid cancer, and to verify the prognostic and clinical significance of HOXC9 and HOXD10. METHODS: Immunohistochemistry was used to determine the expression of HOXC9 and HOXD10 in 98 pairs of papillary thyroid cancer and paracancer tissues. Clinicopathologic data were collected and analyzed to verify the prognostic and clinical significance of HOXC9 and HOXD10. RESULTS: The expression of HOXC9 and HOXD10 decreased in papillary thyroid cancer. The low expression of HOXC9 was associated with Hashimoto's thyroiditis and lymph node metastasis (P<0.05). The low expression of HOXD10 was associated with extrathyroidal extension and lymph node metastasis (P<0.05). The co-expression rates of HOXC9 and HOXD10 was 44.90%. The low expression of both HOXC9 and HOXD10 was associated with lymph node metastasis (P<0.05). CONCLUSIONS: The expression of HOXC9 and HOXD10 was downregulated in papillary thyroid cancer. Low expression of HOXC9 and HOXD10 might be related to the malignancy of papillary thyroid cancer. HOXC9 and HOXD10 may be used as diagnostic and prognostic biomarkers in the future.

Long-term treatment results and prognostic factors of synchronous and metachronous squamous cell carcinoma of head and neck and esophagus.

BACKGROUND: The purpose was to investigate the prognosis of patients with synchronous and metachronous squamous cell carcinoma of head and neck (HNSCC) and esophagus (ESCC), and to evaluate the prognostic factors of these patients. METHODS: A retrospective review was performed on 70 patients with synchronous and metachronous HNSCC and ESCC treated in our institution from January 2005 to December 2016. Kaplan-Meier method and Cox proportional hazard model were used to evaluate overall survival (OS) and determine the prognostic factors associated with survival outcomes. RESULTS: The 1-, 2- and 3-year OS rates were 77.1%, 57.1% and 37.1% with the median survival time for 33.5 months. The univariate analysis results revealed that the patients with early-stage of ESCC, metachronous cancer, and receiving surgery for both cancer had better OS (P=0.003; P=0.035; P=0.002). The multivariate analysis showed that the clinical stage of ESCC and receiving surgery for both cancer or not were the independent prognostic factors for OS. CONCLUSIONS: The multidisciplinary treatment outcome is acceptable, especially for patients with early clinical stage ESCC and with chance to receiving surgery for both cancer.

The extent of lymph node yield in central neck dissection can be affected by preoperative and intraoperative assessment and alter the prognosis of papillary thyroid carcinoma.

BACKGROUND: Lymph node yield (LNY) was implemented in the stratification of papillary thyroid cancer (PTC) patients. The effect of LNY may be related to the extent of surgery. This study aims to identify influencing factors for LNY in central compartment neck dissection (CND). METHODS: Data of 13 712 consecutive PTC patients were analyzed retrospectively. Risk factors for LNY in CND and distribution characteristics of LNY were evaluated. Its relationship with prognosis was studied in another cohort of 136 cases. RESULTS: LNY in therapeutic CND was significantly higher than prophylactic CND (Unilateral: 5.55 ± 3.79 vs 3.41 ± 2.77; Bilateral: 8.90 ± 5.10 vs 6.47 ± 4.17, P < .001). Other independent factors included extranodal extension (ETE), tumor size, and concurrent Hashimoto's thyroiditis. The inconsistency distribution of LNY in bilateral CND was associated with preoperative and intraoperative assessment. Patients with significant difference between major and minor LNY suffered from poorer prognosis (10y-RFS: 58.3% vs 92.0%; HR = 6.719, 95%, P < .0001). CONCLUSIONS: CND surgical procedure, ETE, and Hashimoto's thyroiditis were independent factors of LNY. Inconsistent distribution of LNY was associated with prognosis of bilateral PTC patients. The impact of preoperative and intraoperative assessment on the actual extent of CND can be used to explain the relationship between LNY and PTC prognosis.

Multifocal papillary thyroid cancer in children and adolescents: 12-year experience in a single center.

BACKGROUND: Thyroid cancer is the most common endocrine malignant disease in children and adolescents. There is a trend of more conservative strategies including lobectomy and less radioactive iodine therapy (RAI) in multifocal papillary thyroid cancer (PTC) for its good survival outcome. The aim of our study was to define long-time outcome of a large cohort of multifocal PTC patients less than 20 years old treated at our institution. METHODS: Data were collected from 276 cases who were initially diagnosis of PTC under the age of 20 from January 2006 to December 2015 at Fudan University Shanghai Cancer Center. All patients received total/near total thyroidectomy or lobectomy. Therapeutic central-compartment (level VI) and lateral neck lymph node dissection performed for patients with clinically involved neck nodes. RAI therapy used in selected patients. No patients received chemotherapy or kinase inhibitor therapy. Thyroid-stimulating hormone (TSH) suppression therapy was performed in all patients for at least 5 years. RESULTS: Ninety among 276 were multifocal PTC patients and included in this study. The median follow-up time was 54.28 months, ranging from 6.10 to 141.27 months. Fifteen patients had tumor recurrence during the follow-up. On Kaplan-Meier survival curves, lymphovascular invasion and extrathyroidal extension was associated with a decline in recurrence-free survival. However, there was no difference in recurrence-free survival curves in patients no matter which treatment they had received, either lobectomy or total thyroidectomy, RAI or not. CONCLUSIONS: More conservative strategies including lobectomy and less RAI in multifocal PTC among children and adolescents are safe and effective.

The association between tumor's location and cervical lymph nodes metastasis in papillary thyroid cancer.

BACKGROUND: Papillary thyroid cancer (PTC) has a strong propensity to metastasize to the cervical lymph nodes. Little was known currently about whether tumor's location would influence the risk of lymph node metastasis in PTC. METHODS: The study enrolled PTC patients who underwent primary surgical therapy in our center for small unifocal tumor. The tumor's location was evaluated by ultrasound in three axes, three planes and 3D space. Logistic univariate and multivariate analysis were applied to explore the association between tumors' location and the risk of lymph node metastasis in PTC. Different localization methods of thyroid tumors were evaluated using ROC curve. RESULTS: Totally 1,266 PTC patients were enrolled in this study. Univariate and multivariate analyses showed that gender, age, tumor size and tumor's location (in longitudinal axis, longitudinal sagittal plane, longitudinal coronal plane, sagittal coronal plane and 3D space) was associated with central lymph node dissection (CLND); gender, tumor size and tumor's location (in longitudinal axis, coronal axis, longitudinal sagittal plane, longitudinal coronal plane, sagittal coronal plane and 3D space) was related with lateral lymph node dissection (LLND) (P<0.05). In the ROC curve analysis, the 3D location showed the highest predictive value of lymph node metastasis (C-statistics: 0.724 for CLNM; 0.763 for LLNM). The middle posterior lateral (OR=2.575, P=0.028), inferior anterior central (OR=2.829, P=0.016), inferior posterior lateral (OR=2.759, P=0.039) and isthmus tumors (OR=4.526, P=0.001) were at a higher risk of CLNM, and the middle anterior central tumors (OR=0.102, P=0.015) were related with lower risk of LLNM. CONCLUSIONS: Stereotactic localization showed the highest predictive value of lymph node metastasis. The middle posterior lateral, inferior anterior central, inferior posterior lateral and isthmus tumors were at a higher risk of CLNM when compared to other locations. For such patients, careful preoperative evaluation of nodal status should be done.

Using deep convolutional neural networks for multi-classification of thyroid tumor by histopathology: a large-scale pilot study.

BACKGROUND: To explore whether deep convolutional neural networks (DCNNs) have the potential to improve diagnostic efficiency and increase the level of interobserver agreement in the classification of thyroid nodules in histopathological slides. METHODS: A total of 11,715 fragmented images from 806 patients' original histological images were divided into a training dataset and a test dataset. Inception-ResNet-v2 and VGG-19 were trained using the training dataset and tested using the test dataset to determine the diagnostic efficiencies of different histologic types of thyroid nodules, including normal tissue, adenoma, nodular goiter, papillary thyroid carcinoma (PTC), follicular thyroid carcinoma (FTC), medullary thyroid carcinoma (MTC) and anaplastic thyroid carcinoma (ATC). Misdiagnoses were further analyzed. RESULTS: The total 11,715 fragmented images were divided into a training dataset and a test dataset for each pathology type at a ratio of 5:1. Using the test set, VGG-19 yielded a better average diagnostic accuracy than did Inception-ResNet-v2 (97.34% vs. 94.42%, respectively). The VGG-19 model applied to 7 pathology types showed a fragmentation accuracy of 88.33% for normal tissue, 98.57% for ATC, 98.89% for FTC, 100% for MTC, 97.77% for PTC, 100% for nodular goiter and 92.44% for adenoma. It achieved excellent diagnostic efficiencies for all the malignant types. Normal tissue and adenoma were the most challenging histological types to classify. CONCLUSIONS: The DCNN models, especially VGG-19, achieved satisfactory accuracies on the task of differentiating thyroid tumors by histopathology. Analysis of the misdiagnosed cases revealed that normal tissue and adenoma were the most challenging histological types for the DCNN to differentiate, while all the malignant classifications achieved excellent diagnostic efficiencies. The results indicate that DCNN models may have potential for facilitating histopathologic thyroid disease diagnosis.

Deep convolutional neural network Inception-v3 model for differential diagnosing of lymph node in cytological images: a pilot study.

BACKGROUND: In this study, we exploited the Inception-v3 deep convolutional neural network (DCNN) model to differentiate cervical lymphadenopathy using cytological images. METHODS: A dataset of 80 cases was collected through the fine-needle aspiration (FNA) of enlarged cervical lymph nodes, which consisted of 20 cases of reactive lymphoid hyperplasia, 24 cases of non-Hodgkin's lymphoma (NHL), 16 cases of squamous cell carcinoma (SCC), and 20 cases of adenocarcinoma. The images were cropped into fragmented images and divided into a training dataset and a test dataset. Inception-v3 was trained to make differential diagnoses and then tested. The features of misdiagnosed images were further analysed to discover the features that may influence the diagnostic efficiency of such a DCNN. RESULTS: A total of 742 original images were derived from the cases, from which a total of 7,934 fragmented images were cropped. The classification accuracies for the original images of reactive lymphoid hyperplasia, NHL, SCC and adenocarcinoma were 88.46%, 80.77%, 89.29% and 100%, respectively. The total accuracy on the test dataset was 89.62%. Three fragmented images of reactive lymphoid hyperplasia and three fragmented images of SCC were misclassified as NHL. Three fragmented images of NHL were misclassified as reactive lymphoid hyperplasia, one was misclassified as SCC, and one was misclassified as adenocarcinoma. CONCLUSIONS: In summary, after training with a large dataset, the Inception-v3 DCNN model showed great potential in facilitating the diagnosis of cervical lymphadenopathy using cytological images. Analysis of the misdiagnosed cases revealed that NHL was the most challenging cytology type for DCNN to differentiate.

Metabolic reprogramming and its clinical application in thyroid cancer.

Warburg found that tumor cells exhibit high-level glycolysis, even under aerobic condition, which is known as the 'Warburg effect'. As systemic changes in the entire metabolic network are gradually revealed, it is recognized that metabolic reprogramming has gone far beyond the imagination of Warburg. Metabolic reprogramming involves an active change in cancer cells to adapt to their biological characteristics. Thyroid cancer is a common endocrine malignant tumor whose metabolic characteristics have been studied in recent years. Some drugs targeting tumor metabolism are under clinical trial. This article reviews the metabolic changes and mechanisms in thyroid cancer, aiming to find metabolic-related molecules that could be potential markers to predict prognosis and metabolic pathways, or could serve as therapeutic targets. Our review indicates that knowledge in metabolic alteration has potential contributions in the diagnosis, treatment and prognostic evaluation of thyroid cancer, but further studies are needed for verification as well.

Effects of tumor size on prognosis in differentiated thyroid carcinoma smaller than 2 cm.

Benefits of subdividing small-differentiated thyroid carcinoma (sDTC) by tumor size are controversial. We conducted a meta-analysis to investigate whether tumor size is associated with prognosis of sDTC. PubMed and Web of Science databases were searched from their inception to September 2018. The identified studies according to the inclusion/exclusion criteria were analyzed using fixed/random-effects models. Data were calculated and results of the meta-analysis were expressed as odd ratio (OR). sDTC was classified as S1 (≤1 cm) and S2 (>1 cm and ≤2 cm). A systematic analysis was performed to compare the difference of recurrence, survival and clinicopathological factors between the two subgroups of sDTC (S1 vs. S2). A total of 21 studies published between 2004 and 2017 enrolling 219,291 patients were included. Findings showed that, S2 was associated with higher recurrence risk compared with S1 (OR=1.575, 95% CI=1.428-1.738; P<0.05). There was no statistical difference in survival between S1 and S2, but significant statistical heterogeneity (OR=1.160, 95% CI=0.810-1.662; P=0.448; I2=75.8%). Meta-regression analysis revealed publication year potentially caused the heterogeneity (P<0.05). Comparison of small papillary thyroid carcinoma alone agreed with the results of sDTC. T1b increased the risk of recurrence (OR=1.520; 95% CI=1.072-2.155; P<0.05) and death (OR=1.504; 95% CI 1.353-1.672; P<0.05) compared with T1a. S2 associated with extrathyroidal extension (OR=2.575; 95% CI=1.603-4.135; P<0.05), bilaterality (OR=2.278; 95% CI=1.905-2.723; P<0.05), vascular invasion (OR=4.494; 95% CI=2.812-7.183; P<0.05) and lymph node metastases (OR=1.12; 95% CI=1.10-1.14; P<0.05). Our analysis suggested it is necessary to subdivide sDTC into S1 and S2 owing to their different effects on prognosis, especially recurrence.

Multiple bone brown tumor secondary to primary hyperparathyroidism: a case report and literature review.

Bone brown tumors secondary to primary hyperparathyroidism (PHPT) are rare and only around 2-5% of PHPT patients have multiple bone brown tumor lesions, which are also uncommon in literatures. We found a female patient who got misdiagnosis of multiple malignant bone tumors in our clinical work, she was eventually diagnosed as a brown tumor secondary to hyperparathyroidism. This article records the diagnosis and treatment process, and summarizes similar case reports in the past decade to provide experience in diagnosis and treatment of similar cases that may occur in the future.

NRG1 regulates redox homeostasis via NRF2 in papillary thyroid cancer.

Thyroid cancer is a common endocrine cancer, of which papillary thyroid cancer (PTC) is the most common type. Neuregulin 1 (NRG1), a glycoprotein mediating cell­cell signaling, plays vital roles in cellular activities; however, its role in PTC progression remains poorly understood. In this study, we performed immunohistochemistry in 196 samples from patients and found that NRG1, a potential prognostic marker is highly expressed in PTC compared with adjacent normal tissues. Cell Counting kit­8 (CCK­8) and clone formation assays indicated that NRG1 is essential for PTC cell viability and proliferation, probably by regulating redox homeostasis, which was implied by ROS generation analysis and intracellular GSH activity assay. Western blot analysis and RT­qPCR revealed that NRG1 regulates ERK pathway and the pivotal regulator of cellular redox status, nuclear factor E2­related factor 2 (NRF2), which maintains moderate reactive oxygen species (ROS) levels through a set of antioxidant response element (ARE)­containing genes. The immunohistochemical scoring of 196 PTC samples and the analysis of the data of 490 patients from The Cancer Genome Atlas (TCGA) reveled a positive association between the expression of NRG1 and NRF2. Since the presence of NRG1 regulates redox homeostasis through NRF2, protecting PTC cells from the accumulation of ROS and ROS­induced cell death, NRG1 may thus prove to be a potential therapeutic target in the treatment of thyroid cancer.

Verteporfin inhibits papillary thyroid cancer cells proliferation and cell cycle through ERK1/2 signaling pathway.

Verteporfin, a FDA approved second-generation photosensitizer, has been demonstrated to have anticancer activity in various tumors, but not including papillary thyroid cancer (PTC). In current pre-clinical pilot study, we investigate the effect of verteporfin on proliferation, apoptosis, cell cycle and tumor growth of PTC. Our results indicate verteporfin attenuates cell proliferation, arrests cell cycle in G2/S phase and induces apoptosis of PTC cells. Moreover, treatment of verteporfin dramatically suppresses tumor growth from PTC cells in xenograft mouse model. We further illustrate that exposure to MEK inhibitor U0126 inactivates phosphorylation of ERK1/2 and MEK in verteporfin-treated PTC cells. These data suggest verteporfin exhibits inhibitory effect on PTC cells proliferation and cell cycle partially via ERK1/2 signalling pathway, which strongly encourages the further application of verteporfin in the treatment against PTC.

The Positive Lymph Node Number and Postoperative N-Staging Used to Estimate Survival in Patients with Differentiated Thyroid Cancer: Results from the Surveillance, Epidemiology, and End Results Dataset (1988-2008).

BACKGROUND: Lymph node metastasis is important when evaluating the prognosis of patients with differentiated thyroid cancer (DTC). However, the current N-staging system cannot fully reflect the clinical significance of cervical lymph node metastasis in DTC. In this study, we employed Surveillance, Epidemiology, and End Results (SEER)-registered DTC cases with lymph node metastasis to determine whether the positive lymph node number (PLNN) could be used to improve stratification of patients in terms of survival. METHODS: We used the SEER dataset to identify all DTC patients with at least one positive cervical lymph node who were examined between 1988 and 2008. Multivariable modeling was used to compare cancer-specific survival (CSS) and overall survival (OS) and to calculate different PLNN cutoff points. RESULTS: In total, 14,359 pN + DTC patients identified in the SEER were included. In multivariate Cox regression analysis, the PLNN was significantly associated with both CSS and OS, whereas neither the lymph node ratio (LNR) nor the (numbers of) lymph nodes examined (LNE) were so associated. The highest C-index value (0.933) and the lowest AIC value (9362.687) obtained indicated that the PLNN better predicted the CSS of DTC than did the LNR or LNE. As the p values for both CSS and OS were minimized, and as the PLNN performed best when cases were grouped, PLNN cutoff points of 10 and 3/10 efficiently stratified DTC patients into two and three levels, respectively. Based on the 3/10 trichotomy, the benefits of radioactive iodine (RAI) treatment were evaluated for each group. Such treatment afforded about a 10% survival benefit in patients with more than 10 lymph node metastases. CONCLUSIONS: Compared with the LNR and LNE under different statistical models, PLNN was superior in terms of DTC staging. A cutoff point of 3/10 was optimal for stratifying patients according to prognosis and was of clinical significance in terms of RAI treatment selection.

Aberrant hypermethylation of the HOXD10 gene in papillary thyroid cancer with BRAFV600E mutation.

Epigenetic abnormalities as well as genetic abnormalities may play a vital role in the tumorigenesis of papillary thyroid cancer (PTC). The present study aimed to analyze the function and methylation status of the HOXD10 gene in PTC and aimed to identify relationships between HOXD10 methylation, HOXD10 expression, BRAF mutation and clinicopathological characteristics of PTC. A total of 152 PTC patients were enrolled in the present study. The methylation status of the HOXD10 promoter was analyzed by quantitative methylation-specific polymerase chain reaction (Q-MSP). BRAFV600E mutation status was analyzed by polymerase chain reaction (PCR) followed by DNA sequencing. HOXD10 mRNA expression level was analyzed by real-time polymerase chain reaction (RT-PCR). 5-Aza-2-deoxycytidine (5-Aza) treatment was performed in 4 PTC cell lines to observe the change in HOXD10 expression. Transwell, cell cycle and apoptosis assays were then performed in an HOXD10-overexpressing PTC cell line. Furthermore, we analyzed the associations between HOXD10 methylation, HOXD10 expression, BRAF mutation and clinicopathological characteristics in PTC. Overexpression of HOXD10 suppressed the migration of PTC cells, and promoted cell apoptosis. Q-MSP showed that methylation levels of the HOXD10 promoter were significantly higher in PTC tissues than levels in the adjacent normal thyroid tissues (P=0.02). In addition, expression of HOXD10 was decreased in the PTC cell lines and PTC tissues compared with that noted in the adjacent normal thyroid tissues (P=0.008). However, BRAFV600E mutation was detected in 42.1% of PTC patients enrolled. In addition, the BRAF mutation status was associated with the methylation and expression level of HOXD10 in PTC. We then observed that 5-Aza treatment could revert the expression of HOXD10 in PTC cell lines. Moreover, the hypermethylation of HOXD10 was associated with invasion of the primary tumor and age >45. In conclusion, the HOXD10 gene may act as a tumor suppressor in PTC. The aberrant hypermethylation and decreased expression of the HOXD10 gene were shown in PTC patients, particularly in those with BRAFV600E mutation. The epigenetic suppression of the HOXD10 gene may play a role in the tumorigenesis of PTC, and it is a prospective biomarker for the diagnosis and prognosis of PTC.

BRAF-activated LncRNA functions as a tumor suppressor in papillary thyroid cancer.

Long non-coding RNAs (lncRNAs) participate in cancer cell tumorigenesis, cell cycle control, migration, proliferation, apoptosis, metastasis and drug resistance. The BRAF-activated non-coding RNA (BANCR) functions as both an oncogene and a tumor suppressor. Here, we investigated BANCR's role in papillary thyroid carcinoma (PTC) by assessing BANCR levels in PTC and matched normal thyroid epithelial tissues from 92 patients using qRT-PCR. We also used lentiviral vectors to establish PTC cell lines to investigate the effects of BANCR overexpression on cancer cell proliferation, apoptosis, migration and invasion. Our results indicate BANCR levels are lower in PTC tumor tissues than control tissues. Decreased BANCR levels correlate with tumor size, the presence of multifocal lesions and advanced PTC stage. BANCR overexpression reduced PTC cell proliferation and promoted apoptosis, which inhibited metastasis. It also inactivated ERK1/2 and p38, and this effect was enhanced by treatment with the MEK inhibitor U0126. Finally, BANCR overexpression dramatically inhibited tumor growth from PTC cells in xenograft mouse models. These results suggest BANCR inhibits tumorigenesis in PTC and that BANCR levels may be used as a novel prognostic marker.