Marker For Lymph Node Metastasis Research Articles

Intraductal Prostate Cancer Affinity for Lymphatic-Predominant Metastases Through 18F-DCFPyL‒Prostate-Specific Membrane Antigen‒Positron Emission Tomography/CT Scans in Pretreatment Prostate Cancer Patients.

Intraductal prostate cancer (IDC) is linked to unfavorable oncologic outcomes, marked by distinctive cellular intrinsic pathway changes and intricate immunosuppressive microenvironments that could impact the way cancer spreads. The aim of this study was to determine whether the presence of IDC in prostate biopsy specimens obtained from patients before primary prostate cancer (PCa) treatment is associated with a lymph node metastatic propensity in prostate-specific membrane antigen (PSMA)‒positron emission tomography (PET)/CT. This was a cross-sectional analysis of all PCa patients undergoing a pretreatment 18F-DCFPyL-PSMA-PET/CT between January 1, 2016, and August 2021 at The Princess Margaret Cancer Centre. Outcomes were presence of any metastasis in the overall cohort, presence of lymphatic vs no metastases, and presence of lymphatic vs bone metastasis among patients who underwent PSMA-PET/CT as PCa primary staging. The associations between IDC presence on the prostate biopsy and the study outcomes were evaluated using univariable and multivariable logistic regression analyses. The cohort consisted of 120 patients. IDC and cribriform pattern were observed in 55 (46%) and 48 (40%) prostate biopsies, respectively. Overall, 52 patients (43%) had evidence of metastasis. Presence of IDC on biopsy was associated with increased odds of overall metastasis (odds ratio: 2.47, 95% CI: 1.09-5.61, P = .03). Of the 52 patients with evidence of metastasis, 41 (79%) had evidence of lymphatic metastasis. Presence of IDC on biopsy was associated with significantly increased odds of lymphatic metastasis vs nonmetastases (odds ratio: 3.03, 95% CI: 1.24-7.40, P = .01). The identification of IDC morphology in prostate biopsy specimens has been observed to be significantly linked with lymph node metastasis on 18F-DCFPyL-PET/CT imaging in a PCa pretreatment staging setting. We found that presence of IDC in prostate biopsy appears to be a marker for lymph node metastasis on 18F-DCFPyL-PET/CT.

Leptin as a predictive marker for lymph node metastasis in patients with papillary thyroid cancer

Background Leptin is a neuroendocrine hormone produced from adipose tissue with many vital functions. There is a well-known association between leptin and papillary thyroid cancer (PTC). Preoperative prediction of lymph node metastasis is one of the interesting topics in PTC. This study aimed to determine whether the serum leptin peptide is a significant predictor marker for node metastasis in cases with PTC. Patients and methods This is a prospective comparative study which was conducted at Ain Shams University Hospitals and El-Sheikh Zayed specialized hospital between January 2021 and January 2023. Forty patients with PTC were enrolled in this study and according to postoperative histopathology, we divided them into two groups: group A, 24 cases, those with pathological negative node and group B, 16 cases, those with pathological positive node. Results We had no statistically significant difference in the preoperative demographics data between both groups. Postoperatively, the median serum leptin level and percentage of leptin decrease were significantly higher in in group B. Multivariate logistic regression analysis showed that preoperative serum leptin level, thyroid imaging reporting and data system, multifocality, extra thyroid extension, and size of dominant nodule were significant predictors for node metastasis. There was a significant negative correlation between leptin level and age, while there was a significant positive correlation between leptin level and lymph node metastasis. Conclusion Preoperative serum leptin level is considered a significant predictor marker for lymph node metastasis in PTC.

Sporadic noninvasive medullary thyroid neoplasm: A desmoplasia-negative unifocal nonmetastatic tumor cured by hemithyroidectomy

BackgroundThe absence of primary tumor desmoplasia, a marker of node metastases, on frozen section may help reduce the extent of surgery without compromising the biochemical cure. We aimed to clarify whether hemithyroidectomy with diagnostic ipsilateral central neck dissection can replace total thyroidectomy with routine central neck dissection in patients with sporadic medullary thyroid cancer. MethodsWe retrospectively evaluated data collected from patients who had undergone primary neck surgery for hypercalcitoninemic sporadic medullary thyroid cancer between January 2017 and December 2022 at one institution. ResultsOf the 25 patients we examined, 19 had desmoplasia-negative and 6 desmoplasia-positive primary thyroid tumors on frozen section. The desmoplasia-negative patients had undergone less surgery with fewer nodes removed than the desmoplasia-positive patients (medians of 6 vs 31 nodes, P < .001). The desmoplasia-negative patients had predominantly undergone hemithyroidectomy with ipsilateral central neck dissection. None of the desmoplasia-negative tumors was multifocal (0 of 19 desmoplasia-negative vs 2 of 6 desmoplasia-positive or 0% vs 33%, P = .050) or node-positive (0 of 19 vs 6 of 6 patients or 0% vs 100%; medians of 0 vs 3.5 node metastases; both P < .001). Despite limited surgery, all desmoplasia-negative patients attained and maintained biochemical cure. ConclusionHemithyroidectomy combined with diagnostic ipsilateral central neck dissection is a viable risk-reducing and curative strategy for desmoplasia-negative and node-negative, nonmetastatic unifocal tumors, for which we propose the term sporadic noninvasive medullary thyroid neoplasm (SNMTP).

Increasing C-reactive protein levels in a patient with glioblastoma with lymph node metastasis: a case report

BackgroundGlioblastoma usually recurs locally and extracranial metastases are rare. Most patients with extracranial metastases experience recurrence of the primary intracranial tumor. Lymph node metastases are often detected based on lymphadenopathy or symptoms caused by other metastatic sites.Case presentationHerein, we report a case of glioblastoma with lymph node metastasis in which the patient was asymptomatic but exhibited gradually increasing C-reactive protein levels prior to becoming febrile 9 months after the initial C-reactive protein increase. Diagnosis of lymph node metastasis that was delayed because the patient had a fever of unknown origin, no signs of infection, and the primary intracranial tumor did not recur. Chest computed tomography indicated supraclavicular, mediastinal, and hilar lymphadenopathy, and biopsy identified lymph node metastasis of glioblastoma. This is the fifth reported case of lymph node metastasis without intracranial recurrence.ConclusionsC-reactive protein levels may be a diagnostic marker for lymph node metastasis in patients with glioblastoma. Further evaluation is needed to elucidate the role of CRP in glioblastoma with lymph node metastasis.

TPO as an indicator of lymph node metastasis and recurrence in papillary thyroid carcinoma

The objective of this study was to investigate the expression of thyroid peroxidase (TPO) in papillary thyroid carcinoma (PTC) and to preliminarily investigate its value as a marker of lymph node metastasis and recurrence in patients with PTC. Clinical data of PTC patients and TPO expression were collected from The Cancer Genome Atlas (TCGA) database for analysis. We recruited 230 consecutive PTC patients from the Department of Thyroid Surgery of the First Affiliated Hospital of Kunming Medical University, collected their clinicopathological data, and also performed immunohistochemical analysis of TPO expression on their thyroid specimens to validate the results of bioinformatics analysis. In addition, the construction of protein–protein interaction networks was performed too. Functional enrichment analysis and immuno-infiltration analysis characterized the pathways in which TPO genes may be involved. Data mining based on the TCGA database showed that TPO expression in PTC tissues was significantly lower than in paired normal thyroid tissues. The expression level of TPO in PTC tissues correlated with tumor lymph node metastasis and recurrence. Follow-up data from our center also validated the difference in TPO expression and its relationship with lymph node metastasis in PTC patients. Functional enrichment analysis showed that TPO function was significantly associated with signaling pathways related to amino acid metabolism, gene expression regulation and tumorigenesis. TPO expression was also significantly associated with immune infiltration. Our study showed that reduced TPO expression was significantly associated with lymph node metastasis and recurrence in patients with PTC, and we validated this result in our central cohort. These data suggest that TPO may serve as a prognostic indicator for PTC.

Intratumoral platelet microthrombi in oral squamous cell carcinoma: A marker of lymph node metastasis.

A hypercoagulable state exists in patients with oral squamous cell carcinoma (OSCC), but the role of platelets in the tumour microenvironment has not been explored. This study revealed the status of intratumoral plateletmicrothrombi (PLT-MT) and their clinicopathological relevance and predictive value in OSCC. This study retrospectively evaluated 106 OSCC patients. Tumour and tumour-adjacent tissue specimens were used to stain PLT-MT. Clinicopathological information, patient follow-ups and outcomes and preoperative coagulation and inflammatory hematologic indicators were collected, and their correlation with PLT-MT was analysed. Intratumoral PLT-MT was present in 35 of 106 patients with OSCC who had higher preoperative D-dimer, CRP, FIB and PT levels and lower TT levels. PLT-MT was an independent correlative factor of lymph node metastasis and suggested worse OS in N0 patients. Intratumoral PLT-MT was found in OSCC and was correlated with a hypercoagulable inflammatory state. PLT-MT was an independent marker of lymph node metastasis and showed potential in prognosis prediction.

Abstract 2103: Immunophenotyping of extracellular vesicles isolated from lymphatic exudate identifies CD24 as a marker for lymph node metastases in oral cavity squamous cell carcinoma

Abstract Background: Squamous cell carcinoma of the oral cavity (OSCC) is debilitating and fatal with a high risk of lymph node metastases. One of the key problems is that currently there is no robust molecular biomarker to determine whether the tumor has spread or is at risk of spreading to the regional lymph nodes, the first echelon that OSCC spreads to. Extracellular vesicles (EVs) are functional nanoparticles that are bound by natural protein-lipid bilayers. They are deliberately packaged and proactively secreted by all types of cells and serve as intercellular communication vehicles. Tumors release an excess of EVs which are loaded with the molecular and genetic cargo of the parental tumor cells. Methods: Following ethical approval and written informed consent, pre-surgical plasma as well as matched post-surgical lymphatic drainage fluid were collected from 20 OSCC patients (nodal disease n=6, no node n=14), and control plasma was also collected from healthy donors. EVs were isolated by ultracentrifugation, quantified and characterized by nanoparticle tracking analysis, transmission electron microscopy, and western blot. Immune profiling of 37 EV surface markers was determined using flow cytometry with the MACSPlex Exosome kit. Results: Lymphatic exudate was greatly enriched in EVs in comparison to plasma. Common EV tetraspanins, platelet markers, cancer-related proteins, as well as characteristic markers of immune cells were detected in both plasma and lymphatic exudate EVs based on normalized median fluorescence intensity (nMFI). The presence of OSCC led to an altered composition of blood cell-derived EVs. Elevated levels of CD3, CD25, and CD86, and cancer stem cell marker CD133 were observed in the plasma EVs. However, there were no significant changes in EV markers in plasma between patients with and without lymph node metastasis. Notably, EVs isolated from plasma and lymphatic exudate revealed completely divergent signatures of EV surface markers. Among them, cancer-related protein CD24 was found to be the most highly expressed EV marker in lymphatic exudate from patients with nodal disease (nMFI 1.28±0.50 versus 0.84±0.32, p&amp;lt;0.05) and likewise correlated with advanced disease stages (nMFI 1.17±0.40 versus 0.70±0.27, p&amp;lt;0.05). Conclusions: Plasma-derived EVs are promising non-invasive biomarkers for cancer detection. This study shows that lymphatic exudate is a better source of EV biomarkers due to its high abundance of tumor-related factors directly from draining lymph nodes. In addition, CD24, as reported, is indicative of an immunosuppressive tumor microenvironment and it may serve as a predictor for lymph node metastases and progression of OSCC. These findings shed new light on the understanding of OSCC metastases and provide impetus for consideration of CD24-based liquid biopsy testing and possibly novel immunotherapy. Citation Format: Xinyu Qu, Leanne Lee Leung, Cherrie Wing Kei Ng, Hiu Ching Chan, Zigui Chen, Katie Meehan, Xianhai Zeng, Jason Ying Kuen Chan. Immunophenotyping of extracellular vesicles isolated from lymphatic exudate identifies CD24 as a marker for lymph node metastases in oral cavity squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2103.

ScRNA-seq reveals the correlation between M2 phenotype of tumor-associated macrophages and lymph node metastasis of breast cancer.

The process of lymphatic metastasis was proved to be associated with podoplanin-expressing macrophages in breast cancer (BC). This study aimed to investigate the role of the M2 phenotype of tumor-associated macrophages and mine the key M2 macrophages-related genes for lymph node metastasis in BC. We downloaded the GSE158399 dataset from the Gene Expression Omnibus (GEO) database, which includes transcriptomic profiles of individual cells from primary tumors, negative lymph nodes (NLNs), and positive lymph nodes (PLNs) of breast cancer patients. The cell subsets were identified by clustering analysis after quality control of the scRNA-seq using Seurat. The activation and migration capability of M2 macrophages were evaluated with R package "GSVA". The key M2 macrophages-related genes were screened from the differential expressed genes (DEGs) and M2 macrophages activation and migration gene sets collected from MSigDB database. Our analysis identified three main cell types in primary tumors, NLNs, and PLNs: basal cells, luminal cells, and immune cell subsets. The further cell type classification of immune cell subsets indicated M2 macrophages accumulation in NLs and PLs. The GSVA enrichment scores for activation and migration capability were increased significantly in M2 macrophages from primary tumors than NLNs and PLNs (p-value < 0.001). Seven M2 macrophages activation-related and 15 M2 macrophages migration-related genes were significantly up-regulated in primary tumors than NLNs and PLNs. The proportion and GSVA enrichment scores for activation and migration of M2 macrophages may be potential markers for lymph node metastasis in breast cancer. Our study demonstrated that twenty-two up-regulated mRNA may be possible therapeutic targets for lymph node metastasis in breast cancer.

Histomorphology of immunomodulation and PD1 expression in breast cancer tumor draining lymph nodes

Background: Metastasis in tumor draining lymph nodes (TDLNs) is correlated with poor prognosis in breast cancers. It is associated with local immune suppression, which can be partly due to the higher expression of check point inhibitors in immune cells. The morphological manifestation of the underlying immunomodulation of TDLNs has been less investigated. Here, we present the histomorphological changes and PD1 expression pattern in metastatic and non-metastatic TDLNs in breast cancer patients. Methods: A total of 248 metastatic or non-metastatic TDLNs from 50 breast carcinoma samples were examined histologically and for PD1 expression in the present study. We assessed the immune response in these TDLNs as per histomorphological patterns on H&amp;E stained slides, categorizing them into lymphocyte predominance, germinal center predominance and an un-stimulated pattern. Anti-PD1 immunohistochemistry was performed on all lymph nodes. The results were analyzed using SPSS version 23 and P value &lt;0.05 was considered to be significant. Results: The lymph node metastasis in breast carcinoma was significantly higher at younger age, patients with higher tumor grade and lympho-vascular invasion in the primary tumor. The metastatic lymph nodes showed significantly higher densities of germinal centers with abnormal shapes, as compared to non-metastatic ones. There was significantly higher expression of PD1 in the immune cells of metastatic TDLNs. Conclusion: The identification of PD1 immunohistochemical profile along with histological changes of TDLNs should therefore be considered as a possible prognostic and predictive marker for lymph node metastasis. The patients with higher densities of germinal center with abnormal shape and increased PD1 expression should benefit from immune-check point inhibitor therapy.

Expression of Carboxypeptidase A3 and Tryptase as Markers for Lymph Node Metastasis of Canine Cutaneous Mast Cell Tumors.

Detection of metastatic mast cell tumors (MCTs) in lymph nodes is a critical factor for treatment, prognosis, and clinical management. Presence/absence of mast cells in the lymph nodes cannot be used as a sole parameter to determine metastasis due to the inability to differentiate neoplastic from non-neoplastic/inflammatory mast cells. While cytologic and histopathologic classifications for assessment of metastatic MCTs based on the numbers and distribution of mast cells have been developed, inconsistency between the clinical interpretation of these grading schemes and actual metastatic status occurs. The aim of this study is to identify a novel diagnostic tool to accurately predict overt metastatic mast cell tumors in lymph nodes. We investigated the possibility of using RT-qPCR to detect mRNA expression of mast cell-specific genes in lymph nodes with different stages of MCT metastatic classification. We are able to establish a highly sensitive and discriminating RT-qPCR measuring Carboxy peptidase A3 (CPA3) and tryptase mRNA expression and identify the cut-off values with high sensitivity and specificity for overt metastatic MCTs in lymph nodes. An area of future interest would be to expand our analysis of the extent to which cut-off values for these markers in correctly identifying disease status, as well as predicting clinical outcomes and survival times. This would offer valuable information regarding the practical applicability of this technique and may enable us to improve our standards of detection metastasis, including possibility of molecular analysis of cytologic specimens obtained from suspicious nodes subjected to surgical excision.

Cancer Stem Cell Markers, CD44 and ALDH1, for Assessment of Cancer Risk in OPMDs and Lymph Node Metastasis in Oral Squamous Cell Carcinoma.

Tumour heterogeneity in oral cancer is attributed to the presence of cancer stem cells (CSCs). CSCs are the most migratory and metastatic cellular subpopulation within tumours. Assessment of CSC markers as significant predictors of lymph node metastasis may prove valuable in the clinical setting. Furthermore, analysis of this panel of putative stem cell markers in oral dysplasia may additionally inform of the likelihood for oral potentially malignant disorders (OPMDs) to progress to oral squamous cell carcinoma (OSCC). The present study aims to assess the significance of CSC markers in the progression of OPMDs to OSCC and assessment of lymph node metastasis in OSCC. CD44 and ALDH1 were assessed immunohistochemically in 25 normal, 30 OPMDs, and 24 OSCCs. CD44 is a membranous marker and ALDH1 is a cytoplasmic marker. The immunohistochemical expression of these markers were compared between OPMDs with and without dysplasia, as well as between low-risk and high-risk dysplasias. Similarly, expression was compared between OSCC with and without lymph node metastasis and among grades of OSCC. Positive CD44 expression was seen in all normal mucosal tissues. The expression decreased from normal epithelium to OPMDs but increased in OSCC. CD44 expression was positive in 21 cases of OSCC (87.5%) and reduced from well-differentiated to poorly differentiated OSCC. CD44 staining index was higher in OSCC without lymph node metastasis (3.59) when compared with OSCC with lymph node metastasis (1.33). There was a statistically significant difference observed in the ALDH1 staining index among three groups (p < 0.05), with highest expression seen in OSCC. Within OPMDs, the ALDH1 staining index was statistically higher in OPMDs with dysplasia as compared to OPMDs without dysplasia. Furthermore, the expression was higher in OPMDs with high-risk dysplasia when compared with low-risk dysplasia, but this was not statistically significant (p = 0.82). In conclusion, The CD44 positive population possesses properties of CSCs in head and neck carcinoma, and continuous shedding could be found after CD44 down-regulation. The present study reports differences in ALDH1 expression between OPMDs with and without dysplasia, dysplastic and non-dysplastic epithelia, and low-risk and high-risk dysplasia. These findings may suggest ALDH1 as a specific marker for dysplasia. CD44 demonstrated a difference in staining index in OSCC without lymph node metastasis versus OSCC with lymph node metastasis. These findings may suggest CD44 as a marker for lymph node metastasis. Both proteins may play key roles in the tumorigenicity of CSCs in OPMDs and OSCC.

Nodal metastatic load in papillary thyroid carcinoma. Morphological and molecular analysis with one-step nucleic acid amplification on more than 550 lymph nodes

Background and objectiveThe risk of recurrence in papillary thyroid carcinoma (PTC) is likely related to the amount of tumour in the metastatic lymph node (LN). Therefore, the current TNM classification (N0/N1) make it necessary to find a method to quantify the LN metastasis (LNM). We propose that the quantitative molecular assay One-Step Nucleic-Acid Amplification (OSNA), which measures the number of cytokeratin-19 (CK-19) mRNA copies as a marker of LNM, could play this role. Our objective was to describe the characteristics of the LNs from PTC, and to compare the morphological characteristics that have been claimed as criteria for metastatic burden with OSNA. Patients and methodProspective study of LNs from 42 patients. All of the LNs were measured, weighed and analysed by OSNA and also by imprint cytology. ResultsA total of 573 LNs were included, 187 (32.6%) of them were OSNA-positives. The global consistency between cytology and OSNA was 87.4%. Significant differences were observed in the CK-19 copy number between the LNMs<0.2cm and those >3cm, as well as between those from 0.2 to 3cm with respect to those >3cm, but not between those <0.2cm and those between 0.2 and 3cm. The total tumour load per neck dissection showed no differences based on whether there were ≤5 or >5 LNMs. ConclusionsIn our series the LNMs >3cm show an increased tumour load, but it is unclear if it is necessary to sub-classify the smaller ones as well as the relevance of the number of metastatic nodes according to the cut-off of 5 nodes. We consider that the OSNA analysis avoids the bias of nodal histology and allows for a greater understanding of its real oncological potential.

Expression of Estrogen Receptor- and Progesterone Receptor-Regulating MicroRNAs in Breast Cancer.

In ~70% of breast cancer (BC) cases, estrogen and progesterone receptors (ER and PR) are overexpressed, which can change during tumor progression. Expression changes of these receptors during cancer initiation and progression can be caused by alterations in microRNA (miR, miRNA) expression. To assess the association of BC progression with aberrant expression of miRNAs that target ER and PR mRNAs, we quantified miR-19b, -222, -22, -378a, and -181a in BC samples (n = 174) by real-time PCR. Underexpression of miR-222 and miR-378a in stage T2–T4 BC was characteristic for HER2-overexpressing tumors. In addition, the expression of miR-181a and miR-378a was higher in these tumors than in tumors with a HER2 IHC score of 0 or 1+. In tumors with a Ki-67 index ≥ 14%, all tested miRNAs were underexpressed in BC with a high Allred PR score (6–8). In ER-and-PR–negative tumors, miR-22, miR-222, miR-181a, and miR-378a underexpression was associated with Ki-67 index > 35% (median value). MiR-19b and miR-22 underexpression could be a marker of lymph node metastasis in ER- and/or PR-positive tumors with HER2 IHC score 0. Thus, the association of miR-19b, miR-22, miR-222, miR-378a, and miR-181a levels with BC characteristics is influenced by the status of tumor ER, PR, HER2, and Ki-67.

PAI-1 expression in intratumoral inflammatory infiltrate contributes to lymph node metastasis in oral cancer: A cross-sectional study

IntroductionImmune cells contribute with mediators in the protein expression profile of the tumor microenvironment. Levels of plasminogen activator inhibitor-1 (PAI-1) are elevated in non-malignant inflammatory conditions; however, the association between PAI-1 expression and inflammation remains uncertain in oral squamous cell carcinoma (OSCC). This study aimed to investigate PAI-1 expression in mononuclear inflammatory cell infiltrate in OSCC and its role as a prognostic marker. MethodsSamples were collected from patients with OSCC, treated surgically, and followed for 24 months after the procedure. Thirty-nine tumoral tissue were analyzed using immunohistochemistry. Correlation between protein expression, clinicopathological parameters, and the prognosis was investigated. ResultsPositive PAI-1 expression in mononuclear inflammatory cell infiltrate was significantly associated with lymph node status (p = 0.009) and with the cytoplasmic expression of vascular endothelial growth factor A (VEGFA) (p = 0.028). Multivariate analysis revealed weak PAI-1 expression as an independent marker for lymph node metastases, with approximately 8-fold increased risk compared to strong expression (OR = 8.60; CI = 1.54–48.08; p = 0.014). ConclusionOur results suggest that the strong PAI-1 expression in intratumoral inflammatory infiltrate is an indicator of a better prognosis for patients diagnosed with oral squamous cell carcinoma.

Inhibitor of DNA Binding 2 (ID2): A Novel Marker for Lymph Node Metastasis in Head and Neck Squamous Cell Carcinoma.

Although aggressive invasion and sequential lymph node metastasis (LNM) significantly affect the prognosis of patients with head and neck squamous cell carcinoma (HNSCC), studies on identifying the factors that regulate this process remain scarce. This study found an inhibitor of DNA binding 2 (ID2) as a novel molecule involved in the regulation of invasion and LNM of HNSCC and further verified its functional role. The study examined the translational significance between ID2 expression levels and the presence of LNM as well as the prognosis for 119 patients with HNSCC after treatment. In addition, in vitro and in vivo experiments were performed using ID2 gene-modulated HNSCC cell lines to determine the functional role of ID2 in the invasion and LNM of HNSCC. Elevated levels of ID2 expression were closely associated with the presence of LNM in 119 patients with HNSCC, resulting in a poor prognosis. Overexpression of ID2-induced invasion and LNM of HNSCC cells was observed in vitro and in vivo. By contrast, knockdown of the ID2 gene diminished invasion and LNM of HNSCC cells. In addition, the ID2 expression level increased the expression level of matrix metalloproteinase 1 (MMP1), a molecule downstream to ID2. Furthermore, silencing of MMP1 in ID2-overexpressed HNSCC cells rescued the elevated invasion and LNM capabilities of these cells, suggesting that ID2 enhances invasion and LNM partly via MMP1 activation. In the invasion and LNM of HNSCC, ID2 plays an important role by modulating MMP1 expression, suggesting ID2-MMP1 axis to be a novel alternative therapeutic target for invasion and LNM of HNSCC.

Expression of TMEM16A in Colorectal Cancer and Its Correlation With Clinical and Pathological Parameters.

TMEM16A is a recently identified calcium-activated chloride channel (CaCC) and its overexpression contributes to tumorigenesis and progression in several human malignancies. However, little is known about expression of TMEM16A and its clinical significance in colorectal cancer (CRC). TMEM16A mRNA expression was determined by quantitative real time-PCR (qRT-PCR) in 67 CRC tissues and 24 para-carcinoma tissues. TMEM16A protein expression was performed by immunohistochemistry in 80 CRC tissues. The correlation between TMEM16A expression and clinicopathological parameters, and known genes and proteins involved in CRC was analyzed. The results showed that TMEM16A mRNA expression was frequently detected in 51 CRC tissues (76%), whereas TMEM16A protein expression was determined at a relatively lower frequency (26%). TMEM16A mRNA expression in tumor tissues was higher than its expression in normal para-carcinoma tissues (P < 0.05). TMEM16A mRNA expression was significantly correlated with TNM stage (p = 0.039) and status of lymph node metastasis (p = 0.047). In addition, there was a strong positive correlation between TMEM16A mRNA expression and MSH2 protein. More importantly, TMEM16A protein expression was positively associated with KRAS mutation, and negatively correlated with mutant p53 protein. Logistic regression analysis demonstrated that TMEM16A mRNA expression was an important independent predictive factor of lymph node metastasis (OR = 16.38, CI: 1.91–140.27, p = 0.01). TMEM16A mRNA and protein expression was not significantly related with patient survival. Our findings provide original evidence demonstrating TMEM16A mRNA expression can be a novel predictive marker of lymph node metastasis and TMEM16A protein expression may be an important regulator of tumor proliferation and metastasis in CRC.

PD-L1 Expression Is an Independent Marker for Lymph Node Metastasis in Middle Eastern Endometrial Cancer.

Programmed death ligand 1 (PD-L1) expression in endometrial cancer (EC) tumor cells have been reported in several studies with inconsistent results. Furthermore, there is scarcity of data on the prevalence and prognostic significance of PD-L1 expression in EC from Middle Eastern ethnicity. We aimed to assess PD-L1 expression in a large cohort of Middle Eastern EC and to correlate this with clinico-pathological factors, as well as mismatch repair (MMR) protein status and patients’ outcome. PD-L1 expression was investigated using immunohistochemistry on tissue microarray in an unselected cohort of 440 EC. Kaplan–Meier and logistic regression analysis were used to compare the outcome and prognostic factors. PD-L1 expression in tumor tissue was detected in 18.9% (83/440) EC cases with no impact on survival. When stratified for MMR protein status, PD-L1 expression was similar for both MMR deficient and MMR proficient ECs. However, the expression of PD-L1 in tumor cells was significantly associated with type II (non-endometrioid) histology (p = 0.0005) and lymph node metastasis (p = 0.0172). Multivariate analysis showed PD-L1 expression to be an independent risk factor for lymph node metastasis (odds ratio: 2.94; 95% CI: 1.26–6.84; p = 0.0123). In conclusion, PD-L1 was strongly associated with non-endometrioid EC and was an independent prognostic marker of lymph node metastasis.

Impact of Histological and Molecular Parameters on Prognosis of Oral Squamous Cell Carcinoma: Analysis of 290 Cases.

Background Nodal metastasis is a critical factor in predicting the prognosis of oral squamous cell carcinoma (OSCC). When patients present with a clinically positive neck, the treatment of choice is radical neck dissection. However, management of a clinically negative neck is still a subject of significant controversy. Aim This study was carried out in order to propose a model to predict regional lymph node metastasis of OSCC using histological parameters such as tumour stage, tumour size, pattern of invasion (POI), differentiation of tumour, and host immune response, together with the expression levels of six biomarkers (periostin, HIF-1α, MMP-9, β-catenin, VEGF-C, and EGFR), and, furthermore, to compare the impact of all these parameters on recurrence and 3 yr and 5 yr survival rates. Materials and Method. Histological materials collected from the archives were used to evaluate histological parameters and immunohistochemical profiles. Standard methods were used for immunohistochemistry and for evaluation of results. Data related to recurrence and survival (3 and 5 years) was also recorded. Clinical data was collected from patients' records. Results Male to female ratio was 3 : 1. The commonest site of OSCC was the buccal mucosa, and majority of them were T3 or T4 tumours presented at stage 4. 62.5% of the tumours were well differentiated. Three-year and 5-year survival rates were significantly associated with lymph node metastasis and recurrence. POI was significantly correlated with tumour size, stage, 3-year survival, EGFR, HIF-1α, periostin, and MMP-9 (p < 0.05). Expression of EGFR showed a direct association with metastasis (p < 0.05). Conclusion POI, level of differentiation, and expression of EGFR are independent prognostic markers for lymph node metastasis. Therefore, these parameters may help in treatment planning of a clinically negative neck.

Study of the Relationship Between Serum Amino Acid Metabolism and Lymph Node Metastasis in Patients with Colorectal Cancer.

PurposeLymph node metastasis is one of the important prognostic factors of colorectal cancer, and an important index of individualized treatment. The purpose of this study is to use metabonomics to identify potential molecular markers of lymph node metastasis in colorectal cancer (CRC).Patients and MethodsPeripheral blood samples of 223 CRC patients were collected. The metabolic levels of amino acids and carnitine in peripheral blood of CRC patients, with and without lymph node metastasis, were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS).ResultsThe results show that there were significant differences in the levels of serum amino acids and carnitine between lymph node metastatic patients and lymph node non-metastatic patients. The diagnostic model that was constructed by 9 types of serum metabolites has a high diagnostic ability.ConclusionLC-MS/MS is a detection method that has a broad application in predicting CRC prognosis, individualized treatment, and in studying the mechanism of lymph node metastasis.

157P Search for new markers of thyroid carcinogenesis associated with ER, AR and PR receptor status

In recent years a plethora of new investigating methods have appeared, which allows us to fully and thoroughly observe thyroid diseases. Though, final diagnosis at the pre-operative stage remains to be difficult. Fine-needle aspiration biopsy used for pre-surgical diagnosis doesn't distinguish benign follicular tumors (adenoma) from malignant ones (carcinoma). It is known that thyroid cancer is more common in women (approximately 1: 3), but more aggressive forms of thyroid cancer are common in men. This indicates that estrogen, progesterone and androgen receptors (ER, PR and AR) can play an important role in carcinogenesis of the thyroid gland. Also miRNAs, which role in thyroid cancerogenesis processes is being actively studied, are in interest. However, certain attempts to connect disruptions in receptor expression with disruptions in miRNA expression during thyroid cancerogenesis haven’t been made yet. The expression of the miRNAs and their target-genes was analyzed by RT-PCR in 90 samples of thyroid neoplasms (TN). To find new thyroid cancer markers, we analyzed the expression levels of ER, PR, AR, miRNAs for which these receptors are targets, as well as other target genes of selected miRNAs (ID4, FOXE1, VEGFA, BCL2, ITGB3, SLC24A4, TSHR) in TN. The results has shown that benign TN can be distinguished from malignant ones by expression levels of miRNA-181a, miRNA-205 and their target genes – PR, VEGFA, BCL2, TSHR. Markers, that allow to distinguish follicular variant of papillary thyroid cancer from follicular and papillary carcinomas, have been identified as well. Moreover, it has been shown that miRNA-22 can be marker for lymph node metastases: miRNA level is reduced in patients with metastases. Furthermore, based on the results, it can be assumed that there is a relationship between miRNA-222, miRNA-19b, miRNA-22, miRNA-193b, miRNA-205, miRNA-181a, miRNA-378a levels and ER, PR, AR expression levels during thyroid tumor development. Markers that distinguish between different types of thyroid tumors have been identified (miRNA-181a, miRNA-205, PR, VEGFA, BCL2 and TSHR). The miRNA-22 level was shown to be associated with N stage.