Treatment of anaplastic thyroid cancer with tyrosine kinase inhibitors targeted on the tumor vasculature: initial experience in clinical practice.

Anaplastic thyroid cancer (ATC) is a rarely occurring refractory disease. While recent clinical trials have demonstrated the efficacy of tyrosine kinase inhibitor (TKI) therapy for ATC, evidence is scarce in clinical practice. In this study, we reviewed our initial experiences with TKI treatment in ATC patients with the aim of revealing the efficacy and safety of the same in clinical practice. We retrospectively reviewed our experiences with TKI treatment use in ATC patients diagnosed at our institute from 2014 to 2019. Changes in the patients' neutrophil-to-lymphocyte ratio (NLR) by TKI therapy introduction as well as their clinical factors to indicate the efficacy were examined. Seven patients showed no indication for TKI treatment, while 13 (65%) received treatment. The median duration of TKI treatment was 1.9 months. All patients died, and the overall survival period from diagnosis was 4.7 (95% confidence interval: 2.0-11.5) months. Adverse events ≥Grade 3 were observed commonly (92.3%), and resulted in the termination of TKI treatment in six cases (46.1%). Existence of multiple unfavorable characteristics (higher Prognostic Index) was associated with poor survival. The NLR decreased after the introduction of TKIs and increased again when treatment failed. The response rate to TKI among the ATC patients were approximately 30% in practice. Although the duration of the response was short, several patients demonstrated long survival durations when TKI treatment was provided after successful multidisciplinary treatment to control local disease. Decreases in high NLR values during treatment may suggest the continued effect of TKIs.

Vandetanib versus Cabozantinib in Medullary Thyroid Carcinoma: A Focus on Anti-Angiogenic Effects in Zebrafish Model.

Medullary thyroid carcinoma (MTC) is a tumor deriving from the thyroid C cells. Vandetanib (VAN) and cabozantinib (CAB) are two tyrosine kinase inhibitors targeting REarranged during Transfection (RET) and other kinase receptors and are approved for the treatment of advanced MTC. We aim to compare the in vitro and in vivo anti-tumor activity of VAN and CAB in MTC. The effects of VAN and CAB on viability, cell cycle, and apoptosis of TT and MZ-CRC-1 cells are evaluated in vitro using an MTT assay, DNA flow cytometry with propidium iodide, and Annexin V-FITC/propidium iodide staining, respectively. In vivo, the anti-angiogenic potential of VAN and CAB is evaluated in Tg(fli1a:EGFP)y1 transgenic fluorescent zebrafish embryos by analyzing the effects on the physiological development of the sub-intestinal vein plexus and the tumor-induced angiogenesis after TT and MZ-CRC-1 xenotransplantation. VAN and CAB exert comparable effects on TT and MZ-CRC-1 viability inhibition and cell cycle perturbation, and stimulated apoptosis with a prominent effect by VAN in MZ-CRC-1 and CAB in TT cells. Regarding zebrafish, both drugs inhibit angiogenesis in a dose-dependent manner, in particular CAB shows a more potent anti-angiogenic activity than VAN. To conclude, although VAN and CAB show comparable antiproliferative effects in MTC, the anti-angiogenic activity of CAB appears to be more relevant.

The roles and role-players in thyroid cancer angiogenesis.

Thyroid cancer is the most prevalent endocrine cancer worldwide. Angiogenesis, the formation of new blood vessels, plays a pivotal role in the development and progression of tumors. Over the past years, cancer research has focused on the ability of tumors to induce newly formed blood vessel, because tumor growth and the process of cancer metastasis mainly depends on angiogenesis. Tumor neovascularization occurs following the imbalance between pro-angiogenic and anti-angiogenic factors until the tumor switches to an angiogenic phenotype. A number of signaling factors and receptors that are implicated in the regulation of angiogenesis have been identified and characterized; most notably, the vascular endothelial growth factors (VEGFs) family and their receptors, which are the main pro-angiogenic molecules during early development and in pathological conditions such as cancer. Although thyroid is a highly vascularized organ, angiogenic switch in tumors of this organ leads to the formation of a vast network of blood vessels that favors the dissemination of tumor cells to distant organs and results in deterioration of patient conditions. Accordingly, the identification of key angiogenic biomarkers for thyroid cancer can facilitate diagnosis, prognosis and clinical decision-making and also may help to discover targeting factors for effective cancer therapy as well as monitoring response to therapy. Hence, the main purposes of this review are to summarize the types and mechanisms of angiogenesis emphasizing the prominent factors implicated in thyroid cancer angiogenesis.

Transcription Factor Prospero Homeobox 1 (PROX1) as a Potential Angiogenic Regulator of Follicular Thyroid Cancer Dissemination.

It is well known that Prospero homeobox 1 (PROX1) is a crucial regulator of lymphangiogenesis, that reprograms blood endothelial cells to lymphatic phenotype. However, the role of PROX1 in tumor progression, especially in angiogenesis remains controversial. Herein, we studied the role of PROX1 in angiogenesis in cell lines derived from follicular thyroid cancer (FTC: FTC-133) and squamous cell carcinoma of the thyroid gland (SCT: CGTH-W-1) upon PROX1 knockdown. The genes involved in angiogenesis were selected by RNA-seq, and the impact of PROX1 on vascularization potential was investigated using human umbilical vein endothelial cells (HUVECs) cultured in conditioned medium collected from FTC- or SCT-derived cancer cell lines after PROX1 silencing. The angiogenic phenotype was examined in connection with the analysis of focal adhesion and correlated with fibroblast growth factor 2 (FGF2) levels. Additionally, the expression of selected genes involved in angiogenesis was detected in human FTC tissues. As a result, we demonstrated that PROX1 knockdown resulted in upregulation of factors associated with vascularization, such as metalloproteinases (MMP1 and 3), FGF2, vascular endothelial growth factors C (VEGFC), BAI1 associated protein 2 (BAIAP2), nudix hydrolase 6 (NUDT6), angiopoietin 1 (ANGPT1), and vascular endothelial growth factor receptor 2 (KDR). The observed molecular changes resulted in the enhanced formation of capillary-like structures by HUVECs and upregulated focal adhesion in FTC-133 and CGTH-W-1 cells. The signature of selected angiogenic genes' expression in a series of FTC specimens varied depending on the case. Interestingly, PROX1 and FGF2 showed opposing expression levels in FTC tissues and seven thyroid tumor-derived cell lines. In summary, our data revealed that PROX1 is involved in the spreading of thyroid cancer cells by regulation of angiogenesis.

LncRNA-MALAT1 Promotes Angiogenesis of Thyroid Cancer by Modulating Tumor-Associated Macrophage FGF2 Protein Secretion.

Tumor-associated macrophages (TAMs) in the tumor microenvironment have been associated with enhanced tumor progression. In this study, we investigated the role and molecular mechanisms of MALAT1 in TAMs derived from thyroid cancer. The expression of MALAT1 and FGF2 in thyroid cancer tissues and cells were measured by quantitative real-time PCR and Western blot. TAMs were transfected with indicated constructs. Then the culture medium (CM) from TAMs was harvested for assay. Secreted FGF2 protein levels and TNF-α, IL-12, and IL-10 levels were detected by ELISA. The cell proliferation, migration, and invasion of FTC133 cells were determined with a CCK-8 assay and a Transwell assay, respectively. In addition, HUVEC vasculature formation was measured by matrigel angiogenesis assay. The higher levels of MALAT-1 and FGF2 were observed in thyroid cancer tissues and in thyroid cancer cells compared to that in the control. Besides, in the presence of si-MALAT1, the levels of TNF-α and IL-12 were significantly up-regulated whereas IL-10 was down-regulated in the CM from TAMs. Moreover, down-regulation of MALAT1 in TAMs reduced proliferation, migration, and invasion of FTC133 cells and inhibited angiogenesis. However, overexpression of FGF2 blocked the effects of MALAT1 siRNAs on cell migration, invasion, and angiogenesis. Our results suggest that MALAT1-mediated FGF2 protein secretion from TAMs inhibits inflammatory cytokines release, promotes proliferation, migration, and invasion of FTC133 cells and induces vasculature formation. J. Cell. Biochem. 118: 4821-4830, 2017. © 2017 Wiley Periodicals, Inc.

Recurrent thyroid cancers have more peritumoural lymphatic vasculature than nonrecurrent thyroid cancers.

BACKGROUND: The goal of the study was to evaluate angiogenesis and lymphangiogenesis in differentiated thyroid cancer and recurrences. METHODS: Twenty-seven patients with recurrent differentiated thyroid cancer (20 papillary and seven follicular thyroid carcinomas) and 24 nonrecurrent thyroid cancers were included in this study. Additionally, 24 thyroid adenomas were included as benign controls. All thyroid cancer recurrences were operatively managed, and local recurrences in cervical lymph nodes or cervical soft tissue were histologically confirmed. Altogether, a total of 108 samples were evaluated using CD31 and D2-40 immunohistochemical staining and microscopy. RESULTS: As measured in primary tumours, the median density of CD31-positive vascular structures was 327 vessels (v)/mm(2) for recurrent cancers, 362 v/mm(2) for nonrecurrent cancers and 484 v/mm(2) for thyroid adenomas (P = 0·017). Among the subgroups, the lowest median vascular density of 316 v/mm(2) was found in recurrent papillary cancers and the highest vascular density of 604 v/mm(2) was observed in nonrecurrent follicular cancers (P = 0·018). The median density of D2-40-positive peritumoural lymphatic vessels was 101/mm(2) in recurrent cancers, 56·1/mm(2) in nonrecurrent cancers and 53·9/mm(2) for adenomas (P = 0·015). In the subgroups, peritumoural lymphatic vascular density was 102 v/mm(2) in recurrent papillary cancers and 56·0 v/mm(2) in nonrecurrent papillary cancers (P = 0·044). CONCLUSIONS: Recurrent thyroid cancers expressed less intratumoural microvessels than thyroid adenomas. A high density of peritumoural lymphatic vessels was found in recurrent papillary cancers. High blood vessel density may be a marker for less aggressive tumours, while high peritumoural lymphatic vasculature is a marker for more aggressive and recurrence-prone tumours.

Correlation between maximum intensity and microvessel density for differentiation of malignant from benign thyroid nodules on contrast-enhanced sonography.

OBJECTIVES: The purpose of this study was to retrospectively evaluate contrast-enhanced sonography for differentiation of benign and malignant thyroid nodules by analyzing the correlation between maximum intensity and microvessel density. METHODS: From February 2010 to May 2012, 122 patients (85 female and 37 male; mean age ± SD, 45 ± 9.1 years) with thyroid nodules (62 papillary thyroid carcinomas, 30 nodular goiters, and 30 adenomas) that underwent routine thyroid sonography and were diagnosed by surgery were included in this study. Contrast-enhanced sonography was performed, and enhancement patterns were classified into 3 groups: high, equal, and low enhancement. As a time-intensity curve parameter, the correlation of maximum intensity with CD31 and CD34 microvessel density counts was analyzed. RESULTS: On contrast-enhanced sonography, most patients with papillary thyroid carcinomas showed a heterogeneous low enhancement pattern, whereas most patients with nodular goiters showed an equal enhancement pattern, and patients with adenomas showed a high enhancement pattern. The detection of papillary thyroid carcinomas with low enhancement had sensitivity of 96.8%, specificity of 95.0%, and accuracy of 95.9%. Compared with the papillary thyroid group, the mean microvessel density counts were significantly higher in the nodular goiter and adenoma groups (P< .05). We also found that the maximum intensity was significantly associated with CD31 and CD34 counts (CD31, r = 0.963; P < .01; CD34, r = 0.968; P < .01). CONCLUSIONS: Maximum intensity has a significant relationship with microvessel density. Contrast-enhanced sonography is a practical and convenient means for differentiating benign from malignant thyroid nodules.

S100A4 silencing suppresses proliferation, angiogenesis and invasion of thyroid cancer cells through downregulation of MMP-9 and VEGF.

BACKGROUND AND AIM: It is well documented that S100A4 is upregulated in many cancers and plays a pivotal role in tumor proliferation, invasion, metastasis and angiogenesis. However, the precise role and mechanism S100A4 exerts in the thyroid cancer have not been fully elucidated to date. In the present study, we investigated the effect of S100A4 on proliferation, invasion, metastasis and angiogenesis in thyroid cancer cells. MATERIALS AND METHODS: A plasmid construct was made that expressed full long S100A4 cDNA. The construct was stably transfected into BCPAP and ML-1 thyroid cancer cells (BCPAP/ S100A4 cDNA, ML-1/S100A4 cDNA). S100A4 siRNA was transiently transfected into the DRO cells (DRO/S100A4 siRNA). MMP-9 siRNA or VEGF siRNA was transiently transfected into the BCPAP/ S100A4 cDNA, ML-1/S100A4 cDNA cells (BCPAP/ S100A4 cDNA/VEGF siRNA, ML-1/S100A4 cDNA/ MMP-9 siRNA). RESULTS: We found that the down-regulation of S100A4 by small interfering RNA decreased cell invasion, metastasis, and angiogenesis by using chicken chorioallantoic membrane (CAM), whereas S100A4 overexpression by cDNA transfection led to increased tumor cell invasion, metastasis, and angiogenesis. Consistent with these results, we found that the down-regulation of S100A4 reduced VEGF and MMP-9 expression. Furthermore, Knockdown of MMP-9 by MMP-9 siRNA inhibited cell invasion and metastasis in the BCPAP/S100A4 cDNA, ML-1/S100A4 cDNA cells. Knockdown of VEGF by VEGF siRNA inhibited cell angiogenesis in the BCPAP/ S100A4 cDNA, ML-1/S100A4 cDNA cells.We also found that downregulation of S100A4 by small interfering RNA resulted in enhanced cell growth inhibition and apoptosis, and vice versa. Our data suggest S100A4 could be an effective approach for the regulation of proliferation, invasion and angiogenesis. Downregulation of S100A4 could inhibit angiogenesis, proliferation and invasion by regulating the expression of MMP-9 and VEGF. CONCLUSIONS: Our results provide evidence that the downregulation of S100A4 using RNAi technology may provide an effective tool for thyroid cancer therapy.

Transcription factor Runx2 is a regulator of epithelial-mesenchymal transition and invasion in thyroid carcinomas.

Runx2/Cbfa1 is a member of the Runt-related transcription factor family and is an essential regulator of osteoblast/chondrocyte differentiation. Recently, aberrant expression of Runx2 and its oncogenic functions have been identified in the progression and metastasis of human cancers. In this study, we investigated the expression profile of Runx family genes in normal thyroid tissue, non-neoplastic but abnormal thyroid tissue, various types of thyroid tumors and representative human thyroid carcinoma cell lines. Using reverse transcriptase-PCR and western blotting, we found that Runx2 was consistently upregulated in papillary carcinomas (PCs) and thyroid carcinoma cell lines compared with normal thyroid tissue. With immunohistochemistry, we observed negative or focal immunoreactivity of Runx2 in the nuclei of normal thyroid follicular cells. None of the non-neoplastic thyroid tissues, including Graves' thyroid and adenomatous goiter, had diffuse positivity of Runx2. Expression of Runx2 in benign follicular adenomas varied from negative to diffusely positive. Meanwhile, all malignant thyroid tumors showed some Runx2 immunopositivity. It was diffuse and intense in 83% (19/23) of PCs, 71% (5/7) of follicular carcinomas (FCs) and 40% (4/10) of undifferentiated carcinomas (UCs). In thyroid carcinoma cell lines, the MEK inhibitor U0126 suppressed Runx2, suggesting an association of the MAPK/ERK pathway with Runx2 regulation. Effective silencing of Runx2 by short interfering RNA (siRNA) demonstrated downregulation of EMT-related molecules (SNAI2, SNAI3 and TWIST1), MMP2 and vasculogenic factors (VEGFA and VEGFC) in thyroid carcinoma cells. We also confirmed that Runx2 silencing suppresses thyroid carcinoma cell invasion in transwell assays. In conclusion, this study provides insight into the potential molecular mechanism of thyroid cancer invasion. Our data suggest that enhanced Runx2 is functionally linked to tumor invasion and metastasis of thyroid carcinoma by regulating EMT-related molecules, matrix metalloproteinases and angiogenic/lymphangiogenic factors.

Molecular target based combinational therapeutic approaches in thyroid cancer.

BACKGROUND: Thyroid cancer, as with other types of cancer, is dependent on angiogenesis for its continued growth and development. Interestingly, estrogen has been shown to contribute to thyroid cancer aggressiveness in vitro, which is in full support of the observed increased incidence of thyroid cancer in women over men. Provided that estrogen has been observed to contribute to increased angiogenesis of estrogen responsive breast cancer, it is conceivable to speculate that estrogen also contributes to angiogenesis of estrogen responsive thyroid cancer. METHODS: In this study, three human thyroid cancer cells (B-CPAP, CGTH-W-1, ML-1) were treated with estrogen alone or estrogen and anti-estrogens (fulvestrant and 3,3'-diindolylmethane, a natural dietary compound) for 24 hours. The cell culture media was then added to human umbilical vein endothelial cell (HUVECs) and assayed for angiogenesis associated events. Vascular endothelial growth factor (VEGF) levels were also quantified in the conditioned media so as to evaluate if it is a key player involved in these observations. RESULTS: Conditioned medium from estrogen treated thyroid cancer cells enhanced phenotypical changes (proliferation, migration and tubulogenesis) of endothelial cells typically observed during angiogenesis. These phenotypic changes observed in HUVECs were determined to be modulated by estrogen induced secretion of VEGF by the cancer cells. Lastly, we show that VEGF secretion was inhibited by the anti-estrogens, fulvestrant and 3,3'-diindolylmethane, which resulted in diminished angiogenesis associated events in HUVECs. CONCLUSION: Our data establishes estrogen as being a key regulator of VEGF secretion/expression in thyroid cells which enhances the process of angiogenesis in thyroid cancer. These findings also suggest the clinical utility of anti-estrogens as anti-angiogenic compounds to be used as a therapeutic means to treat thyroid cancer. We also observed that 3,3'-diindolylmethane is a promising naturally occurring anti-estrogen which can be used as a part of therapeutic regimen to treat thyroid cancer.

Blood flow resistance in lesion vessels and normal host vessels evaluated by pulsed Doppler ultrasound.

Blood flow rate in the microcirculation is proportional to the arteriole-venule pressure difference and inversely proportional to the blood flow resistance (BFR). Generally, the BFR studies to date have focused on using invasive methods and were carried out in non-physiological conditions. Moreover, few studies have been concerned with the relationships of BFR between the tumor vessels and the normal host vessels. The present study investigated the BFR in malignant lesion vessels, benign lesion vessels and normal host vessels in physiological conditions using pulsed Doppler ultrasound as a tool and the thyroid as a model, A total of 133 patients with thyroid nodules were included in the study. The results revealed that most of the BFR parameters were higher in malignant lesion vessels than in benign lesion vessels (P=0.001-0.029), as well as lower in normal host vessels than in malignant or benign lesion vessels (P=0.000-0.017); Low to moderate significant positive correlations of BFR between benign lesion vessels and normal host vessels were also found (r=0.358-0.480, P=0.000 for all). Finally and interestingly, low negative correlations between the malignant nodule sizes and some of the BFR parameters were revealed, though these correlations were not statistically significant (r=-0.205--0.261, P=0.108-0.211). Our results suggested that pulsed Doppler ultrasound could be successfully used to measure BFR in physiological conditions and to reveal the BFR relationship between lesion vessels and normal host vessels, as well as the relationship between the lesion sizes and the BFR.

COX-2 expression and tumor angiogenesis in thyroid carcinoma patients among northeast Chinese population-result of a single-center study.

OBJECTIVE: Cyclooxygenase-2 (COX-2), one of the rate-limiting enzymes in the metabolism of arachidonic acid which is reported to be involved in the pathogenesis of many human tumors. As well, Vascular endothelial growth factor (VEGF) is well known to be involved in the infiltration and metastasis of many kinds of cancers. The aim of this study was to further elucidate the clinicopathologic significance of the immunohistochemical expressions of COX-2 and VEGF in thyroid carcinoma. METHODS: Eighty-five patients with thyroid neoplasms were enrolled in our study from December 2003 to January 2010 from the authors' institution retrospectively. Their tumors were examined in the Department of Pathology, the First Bethune Hospital of Jilin University. Immunohistochemistry was performed on paraffin-embedded tissues sections using monoclonal anti-human COX-2 and VEGF antibodies. The tissues were classified into four types: papillary, follicular, medullary and undifferentiated. The patients ranged in age from 23 to 71 years. Breast cancer slides acted as control slides. The immunohistochemical stains were quantified by staining intensity and by the proportion of positively stained cells which were stained brown or yellow. RESULTS: The results were analysed by χ(2) test. COX-2 and VEGF expressions were stronger in thyroid carcinoma than in thyroid adenomas and normal tissues (P<0.01). COX-2 and VEGF expressions in thyroid carcinoma correlated with the tumor type and TNM stage. CONCLUSION: Our results suggest that expression of COX-2 and VEGF may promote angiogenesis of thyroid carcinoma, its infiltration, and metastasis.

Rapamycin inhibits the invasive ability of thyroid cancer cells by down-regulating the expression of VEGF-C in vitro.

The aim of this study was to observe the effects of rapamycin on proliferation, apoptosis and invasion of SW579 in vitro. The proliferation and apoptosis of SW579 cells were detected by methyl thiazolyl tetrazolium and flow cytometry. Transwell assay was used to observe the changes of invasive ability of SW579 cells after being treated with rapamycin. The effects of rapamycin on the expression of mammalian target of rapamycin (mTOR) signalling and vascular endothelial growth factor C (VEGF-C) were observed by Western blot. The inhibition and apoptosis rates increased obviously when the concentration of rapamycin was 20 nm. When the rapamycin concentration was 10 nm, the invasive ability of SW579 cells changed significantly than when it was 5 nm. Our data showed that when the concentrations of rapamycin were over 20 nm, the expression of mTOR and p70S6K decreased significantly, and the expression of PTEN increased notably. There were no remarkable variations observed when we detected the expression of Akt. We found the expression of VEGF-C was high in SW579 cells and decreased slightly when the cells were treated with 5 nm rapamycin. When the concentration of rapamycin was over 5 nm, significant changes were observed. Rapamycin could inhibit the proliferation and induce the apoptosis of human thyroid cancer cells in vitro by mTOR inhibition. No obvious changes observed in the expression of AKT indicated that there might be a feedback loop effect by the mTOR inhibition induced by rapamycin. Rapamycin could inhibit the invasive ability of SW579 cells by down-regulating the expression of VEGF-C.

Role of ultrasound elastography in prediction of malignancy in thyroid nodules.

BACKGROUND: Ultrasonography is considered useful to distinguish between solid and cystic thyroid nodules and to stratify a nodule's risk of cancer as low, medium, or high. Ultrasound (US) elastography has been applied to study the hardness/elasticity of nodules to differentiate malignant from benign lesions. Elastography possibly can solve the dilemma in reaching an accurate diagnosis for the cytologically known as indeterminate nodules. AIM: To evaluate the sensitivity and specificity of US elastography in the diagnosis of thyroid cancer. PATIENTS AND METHODS: This prospective study included 40 patients. The total number of nodules was 46, they were all euthyroid. Laboratory investigations were done including FT3, FT4, and TSH to exclude hot nodules. Neck US, US elastography, and fine-needle aspiration were done to all patients, and US elastography scoring system from 1 to 4 was used. RESULTS: Four out of the 46 studied nodules were malignant. The ROC curve for elastography score (E-score) showed high sensitivity, specificity for the diagnosis of malignant thyroid nodules with a cut-off value of E-score 4 and high significance (p<0.001), the area under curve was 0.92. The sensitivity was 75.0% and specificity was 100%. For E-score more than 2, the sensitivity was 100% and specificity was 85.37%. CONCLUSION: US elastography can be used to increase both the sensitivity and the specificity of US for the detection of malignant thyroid nodules, and so it seems to have great potential as a new tool for the diagnosis of thyroid cancer.

Tumor cells induce COX-2 and mPGES-1 expression in microvascular endothelial cells mainly by means of IL-1 receptor activation.

Prostaglandin (PG) E(2) plays a key role in immune response, tumor progression and metastasis. We previously showed that macrovessel-derived endothelial cells do not produce PGE(2) enzymatically because they do not express the inducible microsomal PGE-synthase-1 (mPGES-1). Nevertheless, differences between macro- and micro-vessel-derived endothelial cells regarding arachidonic acid (AAc) metabolism profile have been reported. The present work was conducted to evaluate the expression of PGE(2)-pathway-related enzymes in human microvascular endothelial cells (HMVEC) in culture and to test the hypothesis that the tumor cell-HMVEC cross talk could increase mPGES-1 expression in HMVEC. We treated HMVEC in culture with human recombinant IL-1ß. IL-1ß induced PGE(2) release and COX-2 and mPGES-1 expression in terms of mRNA and protein, determined by real-time PCR and immunoblotting, respectively. HMVEC constitutively expressed mPGES-2 and cytosolic PGES (cPGES) and the IL-1ß treatment did not modify their expression. PGE(2) synthesized by HMVEC from exogenous AAc was linked to mPGES-1 expression. Immunohistochemistry analysis confirmed mPGES-1 expression in microvessels in vivo. COX-2 and mPGES-1 were also induced in HMVEC by the conditioned medium from two squamous head and neck carcinoma cell lines. Conditioned medium from tumor cell cultures contained several cytokines including the IL-1ß and IL-1α. Tumor cell-induced COX-2 and mPGES-1 in HMVEC was strongly inhibited by the IL-1-receptor antagonist, indicating the important implication of IL-1 in this effect. HMVEC could therefore contribute directly to PGE(2) formed in the tumor. Our findings support the concept that mPGES-1 could be a target for therapeutic intervention in patients with cancer.

Targeted therapy in refractory thyroid cancer: current achievements and limitations.

Thyroid cancer refractory to conventional treatments lacks effective treatment. Targeted therapy is an emerging therapeutic strategy for these cancers, based on preliminary promising results. Tyrosine kinase inhibitors target both specific oncogenic pathways involved in thyroid cancer progression and aspecific mechanisms such as neoangiogenesis. They are generally well tolerated and most adverse events have low-to-moderate severity. Other classes of drugs have been tested, alone or in combination with tyrosine kinase inhibitors, but so far the results have been limited. The aim of this article is to describe the benefits and limitations of innovative drugs that are currently under investigation in patients with refractory thyroid cancer.

Risk Factors for Recurrence of Follicular Thyroid Cancer: A Systematic Review.

Background: In risk assessment of recurrence, papillary thyroid cancer (PTC) and follicular thyroid cancer (FTC) are often grouped together as differentiated thyroid cancer (DTC). However, while risk factors affecting recurrence of PTC are well established, risk factors for recurrence of FTC are not. This systematic review examines risk factors for recurrence of FTC and evaluates their significance. Methods: A systematic search on PubMed and Embase was performed in September 2020, including studies evaluating risk factors for recurrence of FTC. A quality assessment of the enrolled studies was performed. Results: Nine studies (n = 1544 patients) from eight countries were included. The average recurrence rate was 13.6%, and distant metastasis (DM) constituted 64.8% of the recurrent cases. The risk factors examined were sex, age at diagnosis, primary tumor size, degree of invasiveness, focality, positive resection margin, lymph node (LN) metastasis, and DM at diagnosis. Risk factors correlated with recurrence of FTC were age older than 45 years, primary tumor size above 40 mm, widespread invasion, multifocality, positive resection margin, LN metastasis, and DM at diagnosis. Sex was not a statistically significant risk factor. Conclusions: We identified seven risk factors associated with recurrence of FTC. Age and multifocality were found to be of greater impact regarding recurrence risk of FTC compared with PTC. Future research needs to address the impact of different risk factors for recurrence of FTC particularly including age, primary tumor size, angioinvasion, and mutational status.

Is sonographic intra-nodular vascularity a reliable predictor of thyroid malignancy? A UK tertiary centre study.

OBJECTIVE: British Thyroid Association 2014 guidelines emphasised ultrasound assessment of nodules. One ultrasonographic differentiator of debatable relevance is intra-nodular vascularity. This is the first UK study conducted to address this question. METHODS: Ultrasound reports for thyroid surgery patients over 10 years were retrospectively reviewed. Reports documenting 'intra-nodular vascularity or flow' were analysed. Reports identifying peripheral vascularity only or no intra-nodular flow formed the control group. Concordance with final histology was used to determine the odds ratio for malignancy. RESULTS: A total of 306 patients were included, and 119 (38.9 per cent) nodules demonstrated intra-nodular vascularity. Of these, 60 (50.4 per cent) were malignant compared with 42 per cent in the control group. Intra-nodular vascularity was not a statistically significant predictor of malignancy with an odds ratio of 1.39 (p = 0.18, 95 per cent confidence interval, 0.86-2.23). CONCLUSION: Intra-nodular vascularity in isolation was not a reliable predictor of malignancy. This supports other world literature studies. Although intra-nodular flow should not be relied upon in isolation, interpretation in conjunction with other suspicious findings enhances the predictive value.

Angiogenesis-inhibitors for metastatic thyroid cancer.

BACKGROUND: Systemic cytostatic therapies for advanced, metastatic thyroid carcinomas have been poorly effective. Tumor growth and metastasis depend on blood supply and blood vessel formation (angiogenesis). Therefore, inhibition of angiogenesis may represent a promising target for cancer therapy. OBJECTIVES: To evaluate the benefits and risks of angiogenesis-inhibitors for metastatic thyroid cancer when given alone, or in combination with chemotherapy or radiotherapy. SEARCH STRATEGY: We searched The Cochrane Library (2009, Issue 2), MEDLINE (January 2000 to May 2009) and EMBASE (January 2000 to May 2009) databases and abstracts published in annual proceedings for evidence. Attempts were made to identify studies from references in potentially relevant trials. We also searched for ongoing trials. SELECTION CRITERIA: We planned to include randomized controlled trials that compared angiogenesis-inhibitors with other treatments, no treatment, or placebo in participants who had pathologically confirmed advanced thyroid cancer. DATA COLLECTION AND ANALYSIS: Two authors independently evaluated the search results against the selection criteria. Data extraction and risk of bias assessment were not performed because there were no studies that could be included. MAIN RESULTS: We did not identify any studies which met our full inclusion criteria. AUTHORS' CONCLUSIONS: There is currently no reliable evidence available from randomized controlled trials regarding the bene fi ts and harms of the use of angiogenesis-inhibitors for treating advanced thyroid cancer. Several trials are ongoing.

Angiogenesis and angiogenic factor expression in thyroid cancer.

PURPOSE: Increased expression of angiogenic factors and high vascular density characterize tumors with increased invasive and metastatic capability. Anti-vascular endothelial growth factor (VEGF) therapies have shown an important potentiation of chemotherapy and radiotherapy in experimental and clinical studies. The purpose of this study was to investigate whether it could be possible to identify a subgroup of thyroid cancer patients with high angiogenic activity. METHODS: Formalin-fixed paraffin-embedded tissues from 25 papillary and 18 follicular thyroid carcinomas were assessed immunohistochemically for angiogenic activity, i.e. vascular density (VD) and expression of VEGF and basic fibroblast growth factor (bFGF). RESULTS: VD was significantly higher in follicular tumors (p=0.05). Tumors > 4 cm had a significantly higher VD (p=0.001). High VEGF expression was significantly related to high VD (p=0.05). There was no association of bFGF with histological characteristics. CONCLUSIONS: Increased angiogenic activity is a common feature of thyroid carcinomas, particularly in follicular tumors and larger carcinomas. These results support the testing of anti-VEGF therapies in combination with radiotherapy and chemotherapy in advanced thyroid tumors.