Treatment Of Papillary Thyroid Carcinoma Research Articles

Exploration of DNA Methylation-Driven Genes in Papillary Thyroid Carcinoma Based on the Cancer Genome Atlas.

Although the incidence of thyroid carcinoma is reported to be the highest among malignancies of endocrine system, its diagnosis is still unsatisfactory. This study sought to explore the key DNA methylation-driven genes in the development of papillary thyroid carcinoma (PTC) via a bioinformatic analysis based on the Cancer Genome Atlas (TCGA) database and was validated using the Gene Expression Omnibus (GEO) database. The level 3 DNA methylation, mRNA expression, and clinical data of 499 patients with PTC were obtained from the TCGA database. The R package LIMMA, edgeR, and MethylMix were applied to explore the DNA methylation-driven genes in PTC. The ConsensusPathDB software, DAVID, and STRING databases were used for Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses, as well as protein/protein interaction network construction individually. To verify the result, the explored genes were validated using GSE97466 data set retrieved from the GEO database. Fifty-seven (57) methylation-driven genes were detected via MethylMix based on a beta mixture model that compared the DNA methylation state of tumor tissues with that of the normal tissues. Eventually, three genes (TNFRSF1A, CLDN1, and CASP1) were identified to be the most potential biomarkers for the diagnosis or treatment of PTC. These results suggest the crucial roles of TNFRSF1A, CLDN1, and CASP1 in the tumorigenesis of PTC and provide a vital bioinformatic basis for further experimental validations and clinical applications.

Clinical and Sialographic Imaging Features of 131I Radiation-induced Submandibular Gland Sialadenitis

131I radiation-induced sialadenitis is the most frequent complication of 131I treatment for papillary thyroid carcinoma, but little is known about 131I radiation-induced submandibular gland sialadenitis. The purpose of this study was to compare and contrast the clinical and sialographic imaging features of 131I radiation-induced submandibular gland sialadenitis to 131I radiation-induced parotitis. This retrospective cross-sectional study included patients with 131I radiation-induced submandibular gland sialadenitis and parotitis. Clinical records and sialographic image features were evaluated. The predictor variables included age at the time of diagnosis, gender, course of the disease, site of symptoms, and sialographic image grades. The outcome variable was the location of sialadenitis. A student t-test was conducted to analyze the associations between predictor variables and the outcome. The sample was composed of 4 patients with submandibular gland sialadenitis (100% female), 28 with parotitis (85.7% female), and 1 with submandibular gland sialadenitis and parotitis (P<.05). The occurrence of bilateral glands dysfunction was less often in submandibular glands (SMG: 1/4; PG: 19/28). The age and course of disease were not different between submandibular gland sialadenitis and parotitis (Age, SMG: 46.00±13.59years; PG: 50.04±10.71years, P>.05; Course of the disease, SMG: 11.00±16.69months; PG: 6.96±11.18months, P>.05). Radiographically, 7 of 16 patients with parotitis were identified as grade 2 and 9 patients as grade 3. In 3 patients with submandibular gland sialadenitis, 1patient was identified as grade 2 and 2 patients as grade 3. The postoperative pathological results showed that the proliferation of glandular tissue from the hilum of the submandibular gland caused duct stenosis. The results suggest 131I radiation-induced submandibular gland sialadenitis has a lower incidence compared with parotitis and 131I radiation-induced submandibular gland sialadenitis might be related to duct stenosis caused by proliferative glandular tissue after 131I radiation treatment.

Prognostic lncRNA, miRNA, and mRNA Signatures in Papillary Thyroid Carcinoma.

The current focus in the treatment of papillary thyroid carcinoma (PTC) is tumor progression. The aim of this study was to build RNA-based classifiers and develop a comprehensive model to provide progression-free interval (PFI) risk prediction for PTC. The RNAseq data, miRNAseq data, and clinical information of PTC were downloaded from The Cancer Genome Atlas database. Based on the differently expressed RNAs, the least absolute shrinkage and selection operator (LASSO) Cox regression model was utilized to build the RNA-based classifiers for PFI of the patients with PTC. A 6-messenger RNA (mRNA)-based classifier, a 5-long non-coding RNA (lncRNA)-based classifier, and a 4-microRNA (miRNA)-based classifier were constructed to predict the PFI. Patients with high risk based on the constructed RNA-based classifiers had worse prognosis in Kaplan–Meier curve analysis with log-rank test. The areas under the curves of the first, third, and fifth years in the training and testing set were 0.83, 0.82, and 0.82 and 0.67, 0.72, and 0.73 for the 6-mRNA-based classifier, respectively; 0.75, 0.84, and 0.85 and 0.71, 0.67, and 0.71 for the 5-lncRNA-based classifier, respectively; and 0.70, 0.77, and 0.79 and 0.74, 0.67, and 0.66 for the 4-miRNA-based classifier, respectively. The prediction capability of the three RNA-based classifiers was superior to the TNM stage system. Furthermore, a nomogram based on the verified independent prognostic factors was established for the prognostic prediction. The C-index and calibration plots indicated good predictive accuracy of the nomogram. In summary, the 6-mRNA-based classifier and 5-lncRNA-based classifier constructed in this study were independent prognostic factors for PTC.

Clinical value of matrix metalloproteinase-2 and -9 in ultrasound-guided radiofrequency ablation treatment for papillary thyroid carcinoma.

ObjectiveThis study aimed to investigate serum matrix metalloproteinase (MMP)-2 and MMP-9 levels in patients with papillary thyroid carcinoma (PTC).MethodsForty-one patients with PTC undergoing ultrasound-guided radiofrequency ablation (RFA) and 56 controls were included. Serum MMP-2 and MMP-9 levels were determined by enzyme-linked immunosorbent assay before and after surgery. Potential affecting factors were evaluated by logistic regression analysis.ResultsSerum MMP-2 and MMP-9 levels were significantly higher in PTC patients compared with controls, and decreased significantly after surgery. According to receiver operating characteristic curve analysis, diagnostic values for preoperative serum MMP-2 and MMP-9 levels were 82.4% and 86.6%. There was no contrast-agent perfusion in the ablation zone in 88.5% of lesions, and enhancement within or at the lesion edge in 11.4%. The volume reduction at 3 months’ follow-up was >40%. Age, microcalcification, irregular shape, and lesion diameter and number were influencing factors for PTC. Age, and lesion diameter and number were independent risk factors, while calcification and morphology were protective factors.ConclusionSerum MMP-2 and MMP-9 levels have important clinical values for the diagnosis and treatment of PTC by RFA. Preoperative serum MMP-2 and MMP-9 levels, combined with other affecting factors, contribute to disease prognosis.

Mechanism of RET gene mediated EGFR signaling pathway on epithelial-mesenchymal transition, proliferation and apoptosis of papillary thyroid carcinoma cells.

To explore the mechanism of RET gene mediated EGFR signaling pathway on the epithelial-mesenchymal transition (EMT), proliferation and apoptosis of papillary thyroid carcinoma (PTC) cells. PTC TPC-1 cells and human normal thyroid follicular epithelial cells Nthy-ori 3-1 were collected to identify the expression of RET in PTC. Seven groups were divided according to different transfection protocols, including blank group, negative control group, si-RET group, oe-RET group, AG-490 group, NSC 228155 group, and si-RET + NSC 228155 group. After transfection, qRT-PCR was used to identify whether the transfection was successful or not. qRT-PCR and Western blot were performed to detect the mRNA and protein expressions of RET, EGFR signaling pathway related genes, and EMT related genes. Cell migration, invasion, proliferation and apoptosis abilities were further detected by CCK8, cell scratch, transwell and flow cytometry assays, respectively. RET gene was highly expressed in PTC cells (p<0.05). Compared with blank group, oe-RET group and NSC 228155 group had activated EGFR signaling pathway manifesting in the increased expression of EGFR, p-Src, p-FAK, accelerated EMT showing in the increased expression of N-cadherin and Vimentin expression, but decreased E-cadherin expression, increased cell migration, invasion and proliferation, while decreased apoptosis (all p<0.05); si-RET group and AG-490 group had inhibited activation of EGFR signaling pathway, suppressed EMT, decreased cell migration, invasion and proliferation, while increased apoptosis (all p<0.05); while no evident difference was found in si-RET + NSC 228155 group (all p>0.05). Meanwhile, compared with si-RET group, si-RET + NSC 228155 group showed activated EGFR signaling pathway, accelerated EMT, increased abilities of cell migration, invasion and proliferation, while decreased apoptosis (all p<0.05). RET gene is highly expressed in PTC acting as an oncogene. Silencing RET gene expression may inhibit the invasion and promote the apoptosis of PTC cells by inhibiting the activation of EGFR signaling pathway and mediating the process of EMT. It suggests that RET may offer the possibility of a promising therapeutic target for the treatment of PTC on the basis of the explored mechanism.

Clinical analysis of mediastinal lymph node dissection through sternotomy approach in the treatment of papillary thyroid carcinoma with mediastinal lymph node metastasis

Objective: To investigate the rule of mediastinal lymph node metastasis of papillary thyroid carcinoma and the application of therapeutic mediastinal lymph node dissection through the sternotomy approach in the treatment of mediastinal lymph node metastasis of papillary thyroid carcinoma. Methods: All cases of papillary thyroid carcinoma with mediastinal lymph node metastasis treated through sternotomy cooperated by thoracic surgeons and head and neck surgeons from January 2006 to January 2017 in Cancer Hospital of Chinese Academy of Medical Sciences were included in this study. The distribution, metastasis rate, metastasis degree, surgical method, surgical complications and postoperative survival of patients with mediastinal lymph node metastasis were retrospectively analyzed. Results: A total of 31 patients (16 males and 15 females) with papillary thyroid cancer with mediastinal lymph node metastasis, with a median age of 46 (19-65) years, were enrolled in the group. Partial upper sternotomy was used in 28 cases, and total sternotomy was used in 3 cases. The mediastinal lymph nodes of papillary thyroid carcinoma metastasized farthest to the station 6, and the lymph node metastasis rate of each group from high to low was: 2R (61%), 1R (39%), 3A (39%), 1L (16%), 2L (10%), 4R (10%), 5 (3%) and 6 (3%). No metastasis was observed in station 3P, 4L and 7. In addition, the degree of lymph node metastasis at station 2R was the highest, reaching 35% (77/219). Extra-nodal invasion of mediastinal metastatic lymph nodes in thyroid papillary carcinoma is common (23%), easily fuses into masses (23%) and invades peripheral vascular nerves (26%). Up to 29% of blood transfusions are required during or after surgery due to oozing or bleeding (9/31). The 1-, 3-, 5-and 10-year survival rates of patients undergoing surgical treatment were 94%, 94%, 87% and 81%, respectively. Conclusion: Papillary thyroid carcinoma can metastasize to almost all mediastinal lymph nodes except station 3P, 4L and 7. Radical mediastinal lymph node dissection through sternotomy is an effective method for the treatment of mediastinal lymph node metastasis of thyroid papillary carcinoma.

Total thyroidectomy with therapeutic level II-IV neck dissection for papillary thyroid carcinoma: level VI recurrence patterns.

Lateral and central compartments cervical lymph nodes metastases are common among patients with papillary thyroid carcinoma (PTC). Elective level VI neck dissection during thyroidectomy and lateral neck dissection (LND) for the treatment of PTC with lateral compartment lymph node metastases is controversial because of the uncertain benefit in clinical outcomes and increased risks of surgical morbidity. We aimed to determine the potential benefit of elective level VI neck dissection in patients with cN1 papillary thyroid carcinoma (PTC) by investigating the rate and pattern of locoregional recurrence in PTC patients who underwent total thyroidectomy and therapeutic lateral node dissection (LND; levels II-IV) without elective level VI dissection. A retrospective cohort study. Data on demographics, clinical presentation and workup, intraoperative and pathological report, postoperative course, adjuvant therapy, recurrence patterns, and overall survival were retrieved from the medical charts of patients who underwent thyroid surgery in our hospital between January 2006 and December 2017. A total of 1415 thyroidectomies were performed during the study period, of which 802 (56.67%) were for PTC. Of those PTC patients, 228 (28.42%) also underwent LND (levels II-VI) during the same thyroidectomy procedure. Thirty-four (14.91%) of those 228 patients, underwent total thyroidectomy with therapeutic lateral ND II-IV without elective level VI ND. During the follow-up period, five (14.7%) of the latter cohort were diagnosed with recurrence in central neck (level VI) lymph nodes, and four of them (11.7%) were diagnosed with ipsilateral recurrence at level VI. Our results revealed 11.7% rate of clinically significant recurrent disease in ipsilateral level VI which, in our opinion, does not justify routine prophylactic level VI ND dissection when the ipsilateral lateral neck is operated for metastases.

Effect of thyroidectomy on circulating angiogenic cytokines in papillary thyroid carcinoma and benign goiter: Potential for new biomarkers?

BackgroundCirculating angiogenic factors have been associated with clinical outcomes of papillary thyroid carcinoma, although they may also be released in the context of benign multinodular goiter. We sought to investigate the effect of thyroidectomy on the activity and importance of multiple circulating angiogenic factors in papillary thyroid carcinoma and benign multinodular goiter. MethodsBetween May 2015 and December 2016, patients scheduled for total thyroidectomy for papillary thyroid carcinoma or benign multinodular goiter were offered to enroll in this study. Serum levels of angiopoietin-2, fibroblast growth factor-2, hepatocyte growth factor, platelet-derived growth factor-BB, placenta growth factor, heparin-binding epidermal growth factor, and vascular endothelial growth factor-A and -C were collected preoperatively and 2 weeks postsurgery. These levels were measured by enzyme-linked immunosorbent assay and compared with those of 35 healthy control subjects. ResultsSixty patients with a median age of 52 years, 37 of whom were females, were included: 36 had papillary thyroid carcinoma, and 24 had benign multinodular goiter. In both benign multinodular goiter and papillary thyroid carcinoma, preoperative, circulating angiogenic factors levels were increased with respect to controls (P < .0001), and a decrease after total thyroidectomy was observed in the levels of angiopoietin-2 (P < .0001), fibroblast growth factor-2 (P < .0001), hepatocyte growth factor (P < .001), and heparin-binding epidermal growth factor (P < .01 each). Only patients with papillary thyroid carcinomas, however, showed decrease in the postoperative levels of platelet-derived growth factor-BB and vascular endothelial growth factor-A (P = .001 each). ConclusionResults from this study raise the potential for vascular endothelial growth factor-A and platelet-derived growth factor-BB to be used as biomarkers of the effectiveness of treatment of papillary thyroid carcinoma. These results warrant further investigation and may have potential prognostic implications.

MON-506 Assessment of Quality of Life in Persons with Thyroid Cancer and Thyroid Nodules: A Single Center Experience

INTRODUCTION: Despite an excellent prognosis and survival rate, quality of life (QOL) in thyroid cancer is lower than expected. QOL in thyroid cancer survivors is similar to persons with colon and gynecologic cancers, but worse than breast cancer.1 The SF-36 QOL questionnaire has shown to be a valid tool for assessing QOL in thyroid cancer subjects.2 We evaluated QOL in both thyroid nodule and thyroid cancer patients in an institutional cancer registry. METHODS: The Short Form-36 data version 2 (SF-36) QOL data was obtained from single institution prospective bioinformatics thyroid cancer and benign nodule registry. Physical and mental health scores from the SF-36 questionnaire were obtained from both thyroid cancer and nodule patients. Physical and mental QOL responses were scored on a scale from 20-80 and categorized as either “the same or better than the average population,”; “below average,”; or “well below average.” A two sample Wilcoxon rank sum test and a chi-squared test were used to compare QOL between thyroid cancer and nodule subjects, using QOL as a continuous or categorical variable. Univariate descriptive statistics and bivariate analyses were performed using a Wilcoxon Rank Sum and Chi-squared test for categorical QOL data and Kruskal-Wallis for continuous QOL data. RESULTS: We analyzed 321 thyroid cancer and 32 nodule subjects who completed the SF-36 at a single point in time after diagnosis. There was no difference between the groups with regard to sex, age or QOL scores overall. Average age was 43 and 48 years in cancer and nodule groups respectively. Average Physical QOL score was 50.8 (standard deviation SD + 8.8) and 29.6 (SD+ 12.1), respectively (p=0.42). Average Mental QOL score was 48.9 (SD + 9.9) and 48.3 (SD + 8.1) respectively, p=0.16. Physical QOL score was significantly decreased as cancer stage increased when evaluating results as a continuous variable: Stage 1: 51.8 + 8.7; Stage 2: 50.6 +9.5; Stage 3: 48.7 + 7.2; Stage 4: 47.4 + 9.2. Results for mental QOL as a continuous variable by cancer stage showed improvement in reported QOL with increasing cancer stage: Stage 1: 48 (SD+ 9.9); Stage 2: 50.6 (SD + 11.1); Stage 3: 51.1 (SD + 9.13); Stage 4: 53 (SD+ 6.4), p=0.03. CONCLUSIONS: We found no overall difference in physical nor mental QOL between patients with thyroid cancer or benign thyroid nodules. Overall, QOL was in the “same or better than average” for all respondents, but low cancer stage was associated with higher physical and lower mental QOL scores.REFERENCES:1. Barbus, Elena & Peştean, Claudiu & Iulia Larg, Maria & Piciu, Doina. (2016). Quality of life in thyroid cancer patients: A literature review. Clujul Medical. 90. 10.15386/cjmed-7032. Lubitz CC, De Gregorio L, Fingeret AL, et al. Measurement and Variation in Estimation of Quality of Life Effects of Patients Undergoing Treatment for Papillary Thyroid Carcinoma. Thyroid. 2017;27(2):197-206. doi:10.1089/thy.2016.0260

SUN-473 Primary Hyperparathyroidism and Papillary Thyroid Carcinoma, Association or Coincidence?

Background: The Parathyroid glands originate from the Pharyngeal pouches, Inferior from the third and superior from the fourth. Rarely these glands migrate to the thyroid gland, isthmus or thymus and become ectopic. Primary Hyperparathyroidism (PHPT) is a common cause of hypercalcemia in ambulatory patients. It is also more frequent in women and increases with age. Its treatment is often surgical removal of the affected parathyroid gland. We present a case of an ectopic parathyroid adenoma hidden within the thyroid lobe, treated by thyroid lobectomy ultimately leading to the diagnosis and management of Papillary thyroid carcinoma. Case report: A 73-Year-old female with DM, HTN, hyperlipidemia, osteoporosis was referred to the Endocrine clinic for a history of fractures to the right upper and lower extremities after trivial falls, She was subsequently evaluated for metabolic bone disease, noted to have a PTH 78 (n=14-64 pg/ml) with a total serum calcium 9.7 (n=8.6-10.4 mg/dl), 25-OH Vit-D 14 (n=30-100 ng/ml), urinary calcium to creatinine ratio 20 (n=10-320 mg/g). The parathyroid scan showed persistent activity in the area of the inferior margin right thyroid lobe, suggesting a parathyroid adenoma. She had a parathyroidectomy and during the procedure, the parathyroid gland was unable to be visualized. Hence the Right inferior thyroid lobe was removed. The pathology also showed papillary thyroid carcinoma and the patient had a total thyroidectomy. Discussion: The relationship between PHPT and Papillary thyroid carcinoma still remains unclear. Our patient demonstrated a rare circumstance, wherein the presence of a parathyroid adenoma within the thyroid gland has led to early diagnosis and timely treatment of papillary thyroid carcinoma. Few authors reported thyroid malignancy as the most prevalent cancer among patients with PHPT as the primary disorder. While others report concurrence as a coincidental pathology. Nevertheless, we emphasize the importance of surveillance for thyroid pathology in patients with PHPT that can provide better overall patient outcomes.

Surgical Outcomes and Efficacy of Isthmusectomy in Single Isthmic Papillary Thyroid Carcinoma: A Preliminary Retrospective Study

Background and Objectives The optimal extent of surgery needed for isthmic papillary thyroid carcinoma (PTC) is not well-defined, although total thyroidectomy is usually recommended. This study aimed to evaluate the efficacy of isthmusectomy in the surgical treatment of isthmic PTC. Materials and Methods: We retrospectively studied 121 patients who underwent thyroidectomy with or without central neck dissection (CND) for single isthmic PTC from January 2003 to June 2019. We excluded patients who were clinically suspected to have clinically positive cervical lymph node metastasis and gross extrathyroidal extension or to have cancers at sites other than the thyroid isthmus. Patients were divided into three groups according to the extent of thyroidectomy—total thyroidectomy, lobectomy with isthmusectomy (the lobectomy group), and isthmusectomy. Clinical and pathologic characteristics, surgical outcomes, recurrence rate, and survival were analyzed. Results: Of 121 isthmic PTC patients, 28 were men and 93 were women. Total thyroidectomy, lobectomy with isthmusectomy, and isthmusectomy were performed in 70 (57.8%), 40 (33.1%), and 11 (9.1%) patients, respectively. Prophylactic CND was performed in 104 (86%) patients. In a subgroup analysis, patients who underwent total thyroidectomy showed higher postoperative hypoparathyroidism (p < 0.001) than those who underwent lobectomy and isthmusectomy. However, there were no differences in the recurrence rate or survival between the three groups. Conclusion: Thyroid isthmusectomy may be effective in the surgical treatment of small single isthmic PTC. Further studies are necessary to verify this result.

In silico identification of novel non-synonymous variants in metabolic pathway associated target genes of papillary thyroid carcinoma: A way towards future treatment of papillary thyroid carcinoma

Single nucleotide polymorphisms (SNPs) are regarded as one of the most common genetic variations in the human genome, which can affect the structural and functional characteristics of gene/ protein to cause diseases. Thus this study aims to identify the genetic abnormality of papillary thyroid cancer (PTC) which is a complicated stuff to play a role in the evaluation, diagnosis or treatment of PTC patients. This study followed the identified up and down-regulated genes, associated in PTC related metabolic pathways from previous study for single nucleotide polymorphism (SNP) analysis through dbSNP, SIFT, PredictSNP1 and PredictSNP2 which predicted 8 and 6 highly deleterious rsIDS of non-synonymous SNPs for 5 up and 3 down-regulated target genes of PTC respectively, out of which rsIDs rs187536858, rs201950989 of nsSNPs in up-regulated target genes LPAR5, ZMAT3 and nsSNPs with rsIDs rs121913523, rs142070930, rs372232192, rs372333099 in down-regulated genes KIT, GSTM3 respectively were identified as novel nsSNPs from UniProt data for papillary thyroid carcinoma. Hence this study will help to develop a novel biomarker for solving the genetic disruption in PTC patients.

Apatinib Inhibits Cell Proliferation and Induces Autophagy in Human Papillary Thyroid Carcinoma via the PI3K/Akt/mTOR Signaling Pathway

Background: Patients with metastatic radioiodine-refractory papillary thyroid carcinoma (PTC) have limited treatment options and a poor prognosis. There is an urgent need to develop new drugs targeting PTC for clinical application. Apatinib, a novel small-molecule tyrosine kinase inhibitor (TKI), is highly selective for vascular endothelial growth factor receptor-2 (VEGFR2) and exhibits antitumor effects in a variety of solid tumors. Although apatinib has been shown to be safe and efficacious in radioiodine-refractory differentiated thyroid cancer, the mechanism underlying its antitumor effect is unclear. In this report, we explored the effects of apatinib on PTC in vitro and in vivo.Methods: VEGFR2 expression levels were evaluated by immunohistochemistry (IHC), qPCR, and western blotting (WB). The effects of apatinib on cell viability, colony formation, and migration in the Transwell assay were assessed in vitro, and its effect on tumor growth rate was assessed in vivo. In addition, the levels of proteins in signaling pathways were determined by WB. Finally, the autophagy level was assessed by WB, immunofluorescence (IF), and transmission electron microscopy.Results: We found that high VEGFR2 expression is associated with tumor size, T stage, and lymph node metastasis in patients with PTC and that apatinib inhibits PTC cell growth, promotes apoptosis, and induces cell cycle arrest through the PI3K/Akt/mTOR signaling pathway. Moreover, apatinib induces autophagy, and pharmacological inhibition of autophagy or small interfering RNA (siRNA)-mediated targeting of autophagy-associated gene 5 (ATG5) can further increase PTC cell apoptosis.Conclusion: Our data suggest that apatinib can induce apoptosis and autophagy via the PI3K/Akt/mTOR signaling pathway for the treatment of PTC and that autophagy is a potential novel target for future therapy in resistant PTC.

ANGPTL4 Promotes the Proliferation of Papillary Thyroid Cancer via AKT Pathway.

PurposeAlthough papillary thyroid carcinoma (PTC) is associated with a generally favorable prognosis, about 15% of patients present recurrence and distant metastasis in the next decade leading to death. Angiopoietin-like 4 (ANGPTL4) is secreted to circulation and belongs to the angiopoietin-like proteins. The expression of ANGPTL4 was increased in several solid tumor tissues compared to corresponding paracancerous tissues. ANGPTL4 was identified as pro-tumorigenic protein, including stimulating tumor cell growth, promoting tumor metastasis. However, the clinical significance and biological function of ANGPTL4 in PTC is still unclear. Hence, the purpose of this study was to evaluate the role of ANGPTL4 in PTC, investigating the possibility of whether ANGPTL4 could become a novel target for PTC therapy.MethodsWe investigated the expression level of ANGPTL4 and pAKT in PTC and paracancerous tissue by immunohistochemistry. We determined the effect of ANGPTL4 in PTC cell proliferation through cell counting kit-8 (CCK-8) and cell cycle by flow cytometry analysis. Furthermore, the correlation between ANGPTL4 expression levels and PTC cell proliferation from the TCGA data set was analyzed by GSEA. We explored the role of ANGPTL4 on the phosphorylation of AKT and proliferation in PTC cells via overexpression or knockdown assays and AKT inhibitor assay.ResultsIn the present study, we found that ANGPTL4 was highly expressed in both protein and mRNA level in PTC compared with adjacent noncancerous tissues or benign nodule. ANGPTL4 expression increased according to thyroid tumor progression. ANGPTL4 level was positively correlated with the size of PTC. ANGPTL4 increased cell proliferation and decreased cell cycle arrest of PTC. Knockdown of ANGPTL4 inhibited the phosphorylation of AKT. ANGPTL4 regulated PTC cell proliferation through AKT signaling pathway.ConclusionOur findings suggested that ANGPTL4 was increased in PTC compared with adjacent noncancerous tissues, and ANGPTL4 increased cell proliferation and inhibited cell cycle arrest in PTC cells via promoting AKT phosphorylation. The study may provide fundamental information to suggest its suitability as a target for the treatment of PTC.

Effect of actin related protein 2/3 complex subunit 2 gene silencing on the proliferation and apoptosis of papillary thyroid carcinoma TPC-1 cells

Objective To investigate the effect of actin related protein 2/3 complex subunit 2 (ARPC2) gene silencing on the biological characteristics of papillary thyroid carcinoma (PTC) TPC-1 cells through lentivirus-mediated RNA interference. Methods TPC-1 cells infected with nonsense short hairpin RNA (shRNA) sequence lentivirus (shCtrl) was used as the control group. TPC-1 cells infected with ARPC2 shRNA interference sequence lentivirus (shARPC2) was used as the experimental group, in which the expression of ARPC2 gene was specifically interfered. The effects of silencing the expression of ARPC2 gene on the proliferation of TPC-1 cells were detected by using methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, Western blot and colony formation test. Flow cytometry and Western blot were conducted to detect the effect of silencing ARPC2 gene on TPC-1 cells apoptosis and related proteins. Results shARPC2 could efficiently infect TPC-1 cells, and the expression efficiency of green fluorescent protein was over 85%. Compared with the control group, TPC-1 proliferation was inhibited in the experimental group. The ratio of S-phase cells in the experimental group was reduced compared with that in the control group [(14.79±0.21)% vs. (21.13±0.33)%, t = 27.77, P < 0.05]. The ratio of G1 and G2/M-phase cells in the experimental group was increased compared with that in the control group [G1 phase: (67.57±0.08)% vs. (62.06±0.36)%, t=25.56, P < 0.05; G2/M phase: (17.64±0.12)% vs. (16.91±0.17)%, t=6.154, P < 0.05]. Meanwhile, the expressions of cell cycle-related proteins CDK2, CyclinE and CyclinD were reduced in the experimental group. The number of clone formation in the experimental group was less than that in the control group, the difference was statistically significant [(10±2) vs. (161±6), t=9.011, P < 0.05]. In addition, the apoptotic ratio of cells in the experimental group was higher than that in the control group [(8.60±0.77)% vs. (4.08±0.40)%, t=9.011, P < 0.05]. Western blot showed that the expressions of anti-apoptotic factors p21 and bcl-2 were reduced in the experimental group, while the expression of pro-apoptotic factor bax was increased. Conclusion The interference with the expression of ARPC2 regulated by shRNA can inhibit the proliferation, and promote the apoptosis of PTC TPC-1 cells, indicating that ARPC2 may be a possible biological new target for the treatment of PTC. Key words: Thyroid neoplasms; RNA interference; Cell proliferation; Apoptosis

Hypermethylation of the RSK4 promoter associated with BRAF V600E promotes papillary thyroid carcinoma.

Ribosomal S6 kinase 4 (RSK4) is a putative tumor suppressor gene which is inactivated by epigenetic events in a number human malignancies; however, its role in papillary thyroid carcinoma (PTC) remains largely unknown. The aim of this study was to explore the methylation status of the RSK4 promoter in PTC, and to determine its potential role in thyroid carcinogenesis. Reverse transcription‑quantitative PCR (RT‑qPCR) and western blot analyses were performed to examine the RSK4 mRNA and protein levels, respectively. Methylation‑specific PCR (MSP) and bisulfite genomic sequencing (BGS) were used to analyze methylation status of the RSK4 gene. Sanger sequencing was further carried out to detect the BRAF V600E mutation. Cell proliferation assay was finally performed to evaluate the role of hypermethylation in the growth of PTC cells. The association between RSK4 methylation and the clinicopathological characteristics of patients with PTC was assessed. The methylation frequency of the RSK4 promoter in PTC tissues was higher than that in paired paracancerous tissues. Coincidentally, the RSK4 mRNA levels were also downregulated in PTC tissues when compared with the paracancerous counterparts. The hypermethylation of RSK4 was associated with tumor size and lymph node metastasis. Furthermore, the BRAF V600E mutation may influence RSK4 expression and methylation. Moreover, RSK4 hypermethylation was observed in thyroid cancer cell lines, which was consistent with a lack of RSK4 expression. Upon the 5‑Aza‑deoxycytidine treatment of thyroid cancer cells, RSK4 expression was significantly upregulated, while cell proliferation was inhibited. On the whole, the findings of the present study demonstrate that the hypermethylation of the RSK4 promoter may be one of the mechanisms responsible for the poor RSK4 expression in PTC. Thus, these data suggest that RSK4 may serve as a molecular target for the early diagnosis and treatment of PTC.

Herbal Active Ingredients: An Emerging Potential for the Prevention and Treatment of Papillary Thyroid Carcinoma.

Papillary thyroid carcinoma (PTC) is the most common subtype of differentiated thyroid cancers in Asian coastal cities, where the patients have increased risk of potentially high or excessive iodine intake. Given the high metastasis and recurrence of patients with BRAFV600E mutation, the mortality rate of thyroid cancer has recently shown an upward trend. A variety of therapies, including surgery, radiotherapy, and chemotherapy, have been used to treat thyroid cancer, but these therapies still have limitations, including postoperative complications, drug resistance, poor efficacy, or serious side effects. Recent studies have shown the potential of active ingredients derived from herbal medicine in inhibiting PTC via various cell signaling pathways. Some plant-derived compounds, such as apigenin, genistein, and curcumin, are also known to prevent and treat PTC. This article summarizes the recent advances in the structure-functional impact of anti-PTC active ingredients and their effects on PTC cells and tumor microenvironments with an emphasis on their challenges from basic research to clinical practice.

CYTOLOGICAL CHARACTERISTICS OF POSTOPERATIVE METASTASES OF PAPILLARY THYROID CANCER DURING THE DEVELOPMENT OF SECONDARY RADIOIODINE REFRACTORINESS

Radioiodine refractoriness is the main problem in the diagnosis and treatment of papillary thyroid carcinoma. The aim of the study was to investigate the cytological and immunocytochemical changes of thyrocytes in fine-needle aspiration smears of thyroid papillary cancer metastases in the course of the development of secondary radioiodine resistance. A total of 70 postoperative metastases of thyroid papillary cancer (secondary radioiodine refractory metastases, previously responsive to radioiodine, that eventually loses the ability to radioiodine accumulation, radioiodine-avid metastases, primary radioiodine-refractory metastases), immunohistochemical staining of thyroid peroxidase, thyroglobulin, cytokeratin 17 and cytological analysis were performed. Revealing the presence of specific cellular phenotypes and structures in punctuates, a low percentage of thyroid peroxidase and thyroglobulin-positive thyrocytes allows the development of the method of cytological prediction of the radioiodine therapy effectiveness.

Clinical outcomes and risk stratification for papillary thyroid carcinoma presenting with distant metastasis before the era of tyrosine kinase inhibitors.

Radioactive iodine (RAI) therapy has been the mainstay of treatment for papillary thyroid carcinoma (PTC) patients with distant metastasis (DM). Although tyrosine kinase inhibitors (TKIs) were introduced for the treatment of RAI refractory metastatic thyroid carcinoma several years ago, clinical outcomes for PTC patients with DM treated using RAI therapy remain unclear. We retrospectively examined 64 PTC patients (9 men, 55 women) with DM at diagnosis treated using RAI therapy without administration of any kind of chemotherapy or TKIs. Median age of patients was 58 years. Site of DM was the lungs (n = 59), bone (n = 3), and pleural dissemination (n = 2). No patients showed multiple-organ metastases at diagnosis. By the end of the study period, 21 patients had died of PTC. Cause-specific survival rates at 10, 15, and 20 years after initial surgery were 68.2%, 63.6% and 61.1%, respectively. Uni- and multivariate analyses identified age ≥55 years (HR 3.1, p = 0.023), site of DM other than the lungs (HR 13.4, p < 0.0001), and DM with no RAI avidity (HR 5.1, p = 0.0098) as factors independently associated with disease-related death. When analyses were restricted to patients with lung metastasis (n = 59), surgical non-curability was another independent risk factor (HR 5.2, p = 0.0047) in addition to age and RAI avidity. According to risk stratification analysis based on these risk factors, patients with site of DM other than the lungs or with lung metastasis showing ≥2 risk factors among age ≥55 years, DM with no RAI avidity, and surgical non-curability are expected to show higher mortality rates.

The clinical usefulness of chemokine C-X-C Motif Ligand 12 as a diagnostic marker for Papillary Thyroid Carcinoma.

Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer worldwide. It is essential to develop methods for the accurate diagnosis of PTC to avoid unnecessary surgery. The chemokine C-X-C motif ligand 12 (CXCL12) is associated with various cancers. We aimed to investigate the efficacy of CXCL12 in the diagnosis of PTC in fine-needle aspiration (FNA) specimens. We prospectively collected samples from 58 patients who were scheduled for surgical treatment of PTC from 2013 to 2015. Tissue samples of 31 people with benign thyroid conditions were used as controls. Immunocytochemical and immunohistochemical staining for CXCL12 was performed on FNAs and corresponding tissue specimens. B-type Raf kinase (BRAF) V600E mutant protein expression and gene mutation were also analyzed to compare the clinical usefulness. : The mean age of the patients was 49.1 ± 1.4 years and 88.1% were women. Positive CXCL12 staining was observed in 6.5% of benign and in 98.3% of PTC samples; positive BRAF V600E mutant protein expression was found in 19.4% of benign and 93.1% of PTC samples. For the diagnosis of PTC for CXCL12 staining of FNA specimens, the calculated values were 93.1% sensitivity, 90.3% specificity, 94.7% positive predictive value, 87.5% negative predictive value, and 89.1% accuracy. CXCL12 had 100% sensitivity and specificity for the 12 cases of atypia of undetermined significance (AUS) diagnosed in FNA specimens. CXCL12 may be a useful diagnostic tool for PTC, especially when the FNA specimen is classified as AUS.