Lymphovascular Metastasis Research Articles

Interpreting Discordant Monosomy 3 FISH and Chromosomal Microarray Analysis Results in Uveal Melanoma

Uveal melanoma is the most common primary ocular tumor in adults and causes morbidity through lymphovascular metastasis. The presence of monosomy 3 in uveal melanomas is one of the most important prognostic indicators for metastasis. Two major molecular pathology testing modalities used to assess monosomy 3 are fluorescence in situ hybridization (FISH) and chromosomal microarray analysis (CMA). Here, we report two cases of discordant monosomy 3 test results in uveal melanoma enucleation specimens, performed using these molecular pathology tests. The first case is of uveal melanoma from a 51-year-old male that showed no evidence of monosomy 3 when assessed by CMA, but where it was subsequently detected by FISH. The second case is of uveal melanoma from a 49-year-old male that showed monosomy 3 at the limit of detection when assessed by CMA, but where it was not detected by subsequent FISH analysis. These two cases underscore the potential benefits of each testing modality for monosomy 3. Mainly, while CMA may be more sensitive to low levels of monosomy 3, FISH may be best method for small tumors with high levels of adjacent normal ocular tissue. Our cases suggest that both testing methods should be pursued for uveal melanoma, with a single positive result for either test interpreted as indicating the presence of monosomy 3.

Secreted Protein Acidic and Rich in Cysteine (SPARC) and Chemokine’s mRNA expression in Endometrial Cancer versus Normal Endometrium: A Possible Pattern of Haematogenous and Lymphovascular Metastases

Objectives: Secreted Protein Acidic and Rich in Cysteine (SPARC) is a multifunctional glycoprotein, participating in tissue remodeling, morphogenesis and bone mineralization. Furthermore, SPARC controls important mechanisms involved in cancer progression, including angiogenesis regulation. However, in some studies SPARC was found to show tumor suppression while in other a protumorigenic and prometastatic action. In tumor microenvironment some chemokines and their receptors, thanks to their ability to modulate cancer cells migration and proliferation, are involved in the angiogenetic and metastatic process. In this study we compared, in human endometrial cancer tissue (EC) vs normal endometrium counterpart (NE), SPARC with CXCL12, CXCL11, CXCL8, and CXCR7 mRNA expression. Material and Methods: Fresh specimens from 15 patients with EC and corresponding NE were stored at –80°. One mcg of mRNA was reverse-transcribed in cDNA. A Real-Time PCR determined relative cDNA levels of targeted gene mRNA. Results: In EC vs NE, we observed down-regulation of SPARC mRNA in 91% (P<.05), down regulation of CXCL12 mRNA in 91% (P<.001), and down-regulation of CXCR7 mRNA in 91% (P<.001). In EC, SPARC mRNA down-regulation was directly related in 100% of samples to CXCL12 and CXCR7 (P<.001) and in 73% of samples to VEGF (P<.03). Conclusion: In endometrial cancer, under expression of SPARC is directly related to CXCL12 and CXCR7 and this result might be consistent with a SPARC function on tumor progression and invasion mediated by CXCL12 and CXCR7 on blood and lymphatic spread, respectively.

Reconstruction of Immune Microenvironment and Signaling Pathways in Endometrioid Endometrial Adenocarcinoma During Formation of Lymphovascular Space Involvement and Lymph Node Metastasis.

BackgroundThe amplification or mutation of oncogenes and escape from immune surveillance systems promote tumor metastasis. However, subtle changes in the immune microenvironment and signaling pathways are poorly understood during the formation of lymphovascular space involvement (LVSI) and lymph node (LN) metastasis of endometrioid endometrial adenocarcinoma (EEA).Patients and methodsWe detected tumor immunology-related signaling pathways and immunocyte subtypes according to the mRNA levels of 750 oncogenes and genes relating to the tumor microenvironment and immune response using the Nanostring PanCancer IO 360 Panel in 24 paraffin-embedded tissues of EEAs and benign gynecological diseases. Internal reference genes were used for data normalization.ResultsAngiogenesis and immune cell adhesion signaling pathways were activated during LVSI formation of EEA progression. However, during the development of LVSI to LN metastasis, immune system signaling pathways were significantly inhibited, including antigen presentation, cytotoxicity, lymphoid compartment, interferon signaling, and costimulatory signaling pathways. Immune-related genes (CD69, HLA-DOA, ATF3, GBP1, AP2, DTX3L, EGR1, GBP4, TAP1, EIF2AK2, MX1, ISG15, STAT1, and HLA-DRA) were significantly downregulated in EEA with LN metastasis compared to those in EEA with LVSI. Instead, hypoxia, metabolic stress, epigenetic regulation, matrix remodeling, and metastasis signaling pathways were continuously activated in LN metastasis. We also found that neutrophils, macrophages, and mast cells might be involved in LVSI formation and LN metastasis in EEA.ConclusionsEEA with metastatic LNs showed significant immunosuppressive effects. Some oncogenes, matrix remodeling- and hypoxia-related genes, and neutrophil signatures showed higher expression, suggesting their potential as therapeutic targets and offering new immunotherapy strategies in EEA during LN metastasis.

Gene signature characteristic of elevated stromal infiltration and activation is associated with increased risk of hematogenous and lymphatic metastasis in serous ovarian cancer

BackgroundThe clinical significance of hematogenous and lymphatic metastasis in ovarian cancer has been increasingly addressed, as it plays an imperative role in the formation of both intraperitoneal and distant metastases. Our objective is to identify the key molecules and biological processes potentially related to this relatively novel metastatic route in serous ovarian cancer.MethodsSince lymphovascular space invasion (LVSI) is considered as the first step of hematogenous and lymphatic dissemination, we developed a gene signature mainly based on the transcriptome profiles with available information on LVSI status in the Cancer Genome Atlas (TCGA) dataset. We then explored the underlying biological rationale and prognostic value of the identified gene signature using multiple public databases.ResultsWe observe that primary tumors with increased risk of hematogenous and lymphatic metastasis highly express a panel of genes, namely POSTN, LUM, THBS2, COL3A1, COL5A1, COL5A2, FAP1 and FBN1. The identified geneset is characterized by enhanced deposition of extracellular matrix and extensive stromal activation. Mechanistically, both the recruitment and the activation of stromal cells, especially fibroblasts, are closely associated with lymphovascular metastasis. Survival analysis further reveals that the elevated expression of the identified genes correlates to cancer progression and poor prognosis in patients with serous ovarian cancer.ConclusionsOur findings indicate that tumor stroma supports the hematogenous and lymphatic spread of ovarian cancer, increasing tumor invasiveness and ultimately resulting in worse survival. Thus stroma-targeted therapies may improve the clinical outcomes in combination with cytoreductive surgery and chemotherapy.

Needle Tract Implantation Following Fine-Needle Aspiration of Thyroid Cancer

BackgroundNeedle tract implantation (NTI) can occur after thyroid fine-needle aspiration cytology (FNAC). To date, no studies enrolling a large number of patients and studying the cumulative incidence (CI) of NTI have been published. This study analyzed the clinicopathological features of NTI and calculated the CI.MethodsA total of 11,745 patients who underwent first FNAC, resection for thyroid carcinomas and related lymph node metastases, and postoperative ultrasonography in Kuma Hospital between 2006 and 2017 were evaluated. NTI was defined as macroscopically visible and/or sonographically detected recurrent tumors at the sites of the previous aspiration. Lymphovascular invasion and/or nodal metastasis were excluded.ResultsWe found 22 NTI-THY (originated from aspiration of thyroid tumors) and 8 NTI-LN (originating from lymph nodes) cases with CI of 0.15% and 0.37%, and 0.37% and 0.58% at 5 and 10 years after FNAC, respectively. A large percentage (53%) of cases are presented as more aggressive histologic subtypes than their original tumors. Of these NTI patients, 4 (12.1%) died of disease and 10 (33.3%) developed distant metastases but remained alive with stable disease, three (9.1%) developed local recurrences, and the remaining 15 (10%) were free of recurrence.ConclusionsWe demonstrated for the first time the CI and detailed clinicopathological features of thyroid NTI associated with possible histological transformation. Despite the low CI, the risk of death and distant metastases increase manifold in NTI patients than that in patients without recurrence. Therefore, NTI could be an important prognostic variable and impact patient survival.

Integrin-Mediated Macrophage Adhesion Promotes Lymphovascular Dissemination in Breast Cancer

SummaryLymphatic vasculature is crucial for metastasis in triple-negative breast cancer (TNBC); however, cellular and molecular drivers controlling lymphovascular metastasis are poorly understood. We define a macrophage-dependent signaling cascade that facilitates metastasis through lymphovascular remodeling. TNBC cells instigate mRNA changes in macrophages, resulting in β4 integrin-dependent adhesion to the lymphovasculature. β4 integrin retains macrophages proximal to lymphatic endothelial cells (LECs), where release of TGF-β1 drives LEC contraction via RhoA activation. Macrophages promote gross architectural changes to lymphovasculature by increasing dilation, hyperpermeability, and disorganization. TGF-β1 drives β4 integrin clustering at the macrophage plasma membrane, further promoting macrophage adhesion and demonstrating the dual functionality of TGF-β1 signaling in this context. β4 integrin-expressing macrophages were identified in human breast tumors, and a combination of vascular-remodeling macrophage gene signature and TGF-β signaling scores correlates with metastasis. We postulate that future clinical strategies for patients with TNBC should target crosstalk between β4 integrin and TGF-β1.

Papillary Carcinoma of the Breast Among Omani Women: A Case Series and a Literature Review

Breast cancer is the most common cause of death from cancer in women worldwide. In 2012, an estimated of 100,000 cases of invasive breast cancer were diagnosed in the United States. The histopathology type is predominantly ductal in about 70–80% of patients followed by invasive lobular carcinoma in 5–15% of case. Invasive papillary carcinomas of the breast are rare, accounting for less than 1-2% of invasive breast cancers. Occasionally the disease is found in men as well. Papillary carcinoma most frequently occurs in older and post-menopausal women. Triple assessment is essential to reach the diagnosis. The typical sonographic appearance of IPC is of hypoechoic area with soft tissue echoes emerging from the wall of the cyst. Invasive micropapillary carcinoma is a luminal-type breast cancer with a propensity for lymphovascular invasion and regional lymph-node metastasis. Intracystic papillary breast cancer (IPC) is best managed in the context of a multidisciplinary team. Surgical excision of the lump with margins in excess of 2 mm is considered satisfactory. Many studies suggest that papillary carcinoma may have a better prognosis than invasive ductal carcinoma (IDC). However, the little available information about papillary carcinoma of the breast underscores the need for further management related studies.

Demographic, clinicopathologic and survival differences among left sided versus right sided metastatic colorectal adenocarcinoma.

e15068 Background: Clinical outcomes may differ between left versus right sided (CA) independent of stage. We utilized a large database to identify real world differences in characteristics and overall survival (OS) of left versus right sided CA. Methods:We used the National Cancer Database (NCDB) to identify 36,271 patients with metastatic (stage IV) CA diagnosed between the years 2010 to 2012. Right-sided CA was defined as CA of the cecum, ascending colon or hepatic flexure, and left-sided CA was defined as CA of splenic flexure, descending colon, sigmoid colon and rectum. We compared demographic and pathologic features (tumor size, grade, lymphovascular invasion, metastatic site) as well as OS of CA based on laterality. Results:36% of metastatic CAs were right sided. Left versus right sided CAs were more likely to be diagnosed in older patients (median age 68 vs. 62 yr; p<0.01), females (52% vs. 43%; p<0.01), and African Americans (17% vs. 13%; p <0.01). Left-sided CAs were more likely to be >5 cm (58% vs. 54%, p<0.01), grade 3 or 4 (35% vs. 22%; p<0.01), have lymphovascular invasion (62% vs. 50%; p <0.01) and liver metastasis (72% vs 67%; p<0.01) and less likely to have lung (20% vs. 27%;p <0.01) or bone metastasis (5% vs. 6%; p value <0.01). The median OS was lower in right versus left-sided CAs(12 vs. 20 months; p <0.01) Conclusions:Our study suggests important differences in metastatic CAs based on laterality. Patients with right-sided CAs are more likely to be older, females and African Americans, present with larger tumor, lymphovascular invasion and differ in sites of metastases. Future studies should assess any difference in molecular characteristics and tumor biology based on laterality. [Table: see text]

Could pre-operative NE-NBI finding be useful to determine accurate surgical margins in undifferentiated-type early gastric cancer patients?

Early gastric cancer (EGC) is defined as being mucosal or submucosal invasion, and is curative because of the low risk of lymphovascular metastasis (1).

Brain derived neutrophic factor (BDNF) coordinates lympho-vascular metastasis through a fibroblast-governed paracrine axis in the tumor microenvironment.

It has long been known that the tumor microenvironment contributes to the proliferation and survival of neoplasms through the constant interaction with the stromal and immune compartments. In this investigation, we explored the role of cancer-associated fibroblasts (CAFs) in the regulation of the tumor microenvironment in head and neck squamous cell carcinoma (HNSCC) though a complex intercellular BDNF-TrkB signaling system. Our studies show that conditioned media derived from patient-derived CAFs promoted HNSCC cell proliferation, in vitro cell migration, cell invasion and chemotherapy resistance, compared to normal fibroblasts. Furthermore, examination of the in vivo impact of CAF pathophysiology in the tumor microenvironment in animal xenograft models revealed that HNSCC cell lines in combination with CAFs promoted tumor growth and increased incidence of lymphovascular metastasis as compared to injection of tumor cells or CAF cells alone. Using pharmacological and genetic alterations, we mechanistically demonstrate the critical importance of BDNF-TrkB signaling in the tumor microenvironment. These investigations further support the rationale for BDNF/TRKB targeted therapy against in the treatment of HNSCC.

Inhibition of NF-kappa B pathway leads to deregulation of epithelial–mesenchymal transition and neural invasion in pancreatic cancer

NF-κB has an essential role in the initiation and progression of pancreatic cancer and specifically mediates the induction of epithelial–mesenchymal transition and invasiveness. In this study, we demonstrate the importance of activated NF-κB signaling in EMT induction, lymphovascular metastasis, and neural invasion. Modulation of NF-κB activity was accomplished through the specific NF-κB inhibitor (BAY 11-7085), triptolide, and Minnelide treatment, as well as overexpression of IKBα repressor and IKK activator plasmids. In the classical lymphovascular metastatic cascade, inhibition of NF-κB decreased the expression of several EMT transcription factors (SNAI1, SNAI2, and ZEB1) and mesenchymal markers (VIM and CDH2) and decreased in vitro invasion, which was rescued by IKK activation. This was further demonstrated in vivo via BAY 11-7085 treatment in a orthotopic model of pancreatic cancer. In vivo NF-κB inhibition decreased tumor volume; decreased tumor EMT gene expression, while restoring cell–cell junctions; and decreasing overall metastasis. Furthermore, we demonstrate the importance of active NF-κB signaling in neural invasion. Triptolide treatment inhibits Nerve Growth Factor (NGF) mediated, neural-tumor co-culture in vitro invasion, and dorsal root ganglia (DRG) neural outgrowth through a disruption in tumor-neural cross talk. In vivo, Minnelide treatment decreased neurotrophin expression, nerve density, and sciatic nerve invasion. Taken together, this study demonstrates the importance of NF-κB signaling in the progression of pancreatic cancer through the modulation of EMT induction, lymphovascular invasion, and neural invasion.

Diffuse Gastric Carcinoma Undergoes Characteristic Phenotypic Changes in the Intravascular Environment: Evidence for a Reversal of the Epithelial-Mesenchymal Transition in Lymphovascular Metastasis.

This article reports differences between the properties of extravascular carcinoma, which generally forms the vast bulk of a tumor, and those of intravascular carcinoma, at both primary and metastatic lymph node sites. In a morphological and immunohistochemical study of 19 diffuse gastric adenocarcinomas, we report that in comparison to extravascular carcinoma, the intravascular tumor compartment showed frequent and profound phenotypic change, including increased tumor cell cohesion, differentiation and cadherin/catenin expression. For example, greatest cohesion was seen at the intravascular site in 78% ( P = .00006) of primary cancers and in 84% ( P = .000015) of their lymph node metastases. Pan cadherin showed a statistically significant increase at the intravascular metastatic site ( P = .031). We suggest that this change from an extravascular isolated cell phenotype to an intravascular cohesive phenotype represents reversal of the epithelial to mesenchymal transition. Since this proposed reversal of epithelial to mesenchymal transition in intravascular carcinoma is frequently conspicuous in routine histological sections of many types of cancer, as our previous publications have indicated, this process is likely to have widespread significance for the biology of metastasis.

Coupling of LETM1 up-regulation with oxidative phosphorylation and platelet-derived growth factor receptor signaling via YAP1 transactivation.

Persistent cellular proliferation and metabolic reprogramming are essential processes in carcinogenesis. Here, we performed Gene Set Enrichment Analysis (GSEA) and found that that LETM1, a mitochondrial calcium transporter, is associated with cellular growth signals such as platelet-derived growth factor (PDGF) receptor signaling and insulin signaling pathways. These results were then verified by qRT-PCR and immnunoblotting. Mechanistically, up-regulation of LETM1 induced YAP1 nuclear accumulation, increasing the expression of PDGFB, PDGFRB and THBS4. Consistent with this, LETM1 silencing caused loss of YAP1 nuclear signal, decreasing the expression of PDGFB, PDGFRB and THBS4. Immunohistochemical staining consistently indicated a positive association between LETM1 up-regulation, YAP1 nuclear localization and high PDGFB expression. In clinical data analysis, LETM1 up-regulation in thyroid cancer was found to be related to aggressive tumor features such as lymphovascular invasion (LVI, P < 0.001) and lymph node metastasis (LNM, P = 0.011). Multivariate analysis demonstrated that LETM1 up-regulation increases the risk of LVI and LNM (OR = 3.455, 95% CI = 1.537–7.766 and OR = 3.043, 95% CI = 1.282–7.225, respectively). Collectively, these data suggest that up-regulation of LETM1 induces sustained activation of proliferative signaling pathways, such as PDGF signal pathway by AKT induced YAP1 transactivation, resulting in aggressive thyroid cancer phenotypes.

Prognostic Significance of Lymphatic Vessel Density Detected by D2-40 and Its Relation to Claudin-4 Expression in Prostatic Adenocarcinoma.

Background Lymphovascular invasion is an important pathway of metastatic spread and regional lymph node metastasis is the major prognostic factor in prostatic adenocarcinoma. D2-40 is used to identify the lymphatic vessels and to assess the lymphatic vessel density (LVD). Expression of claudin-4 may be related to invasion and progression of carcinoma cells in several primary tumors. Aim To evaluate intra- and peritumoral LVD through immunohistochemical expression of D2-40 in relation to claudin-4 expression and clinicopathological parameters in prostatic adenocarcinoma. Materials and Methods Immunohistochemical staining procedure was performed on 53 paraffin-embedded blocks of radical prostatectomy specimens for prostatic adenocarcinoma using anti D2-40 and claudin-4 antibodies. Sections were evaluated for mean LVD in intratumoral and peritumoral tissues assessed by D2-40 expression. Results LVD in intratumoral tissues was significantly lower compared with peritumoral areas (P = .0001). Peritumoral mean LVD was significantly higher in cases with lymphovascular invasion (P = .041) and in cases with positive lymph node metastasis (P = .003) than intratumoral mean LVD. High claudin-4 expression was significantly correlated with high tumor grade (P = .0001), lymphovascular invasion (P = .006), and positive lymph node metastasis (P = .004). High claudin-4 expression was significantly associated with increased mean LVD in peritumoral tissues. Conclusion Increased peritumoral mean LVD in prostatic adenocarcinoma is associated with lymphovascular invasion and positive lymph node metastasis. High claudin-4 expression is associated with high tumor grade, lymphocascular invasion, positive lymph node metastasis, and high mean peritumoral LVD suggesting that D2-40 and claudin-4 may represent different mechanisms of lymphatic vessel invasion with both biomarkers is related to poor prognosis.

P-069 Risk Factors of Submucosal or Lymphovascular Metastasis during resection of the Early Gastric Cancer by Endoscopic Submucosal Dissection

P-069 Risk Factors of Submucosal or Lymphovascular Metastasis during resection of the Early Gastric Cancer by Endoscopic Submucosal Dissection

Abstract P1-07-27: Neutrophil elastase modulates breast cancer progression by fostering collective cell detachment and tumor emboli dissemination

Abstract Proteases constitute a large family of enzymes involved in several processes, including degradation and remodeling of the extracellular matrix to drive dissemination of cancer cells into adjacent tissue. Within this large family, the serine protease neutrophil elastase (NE) has been proven clinically meaningful in breast cancer. Indeed, high levels of NE correlate with poor outcome and endocrine resistance in breast cancer patients. In an experimental model it has been demonstrated that estrogen receptor positive (ER+) MCF7 breast cancer cells grow in suspension as 3D-spheroids in NE-addicted medium, thus resembling the micropapillae of a human breast cancer characterized by high propensity to metastasize, i.e. the micropapillary carcinoma. We hypothesized that NE may produce disarrangement of tumor cell adhesion to the substrate fostering neoplastic lymphovascular invasion (LVI) and metastasis. ER+/E-Cadherin (E-CAD)+/HER2- (MCF7, T-47D, ZR-75-1), ER+/E-CAD+/HER2+ (BT-474), ER-/E-CAD-/HER2+ (SK-BR-3) and ER-/E-CAD-/HER2- (MDA-MB-231) cells were grown with serine proteases (NE, cathepsin-G), hyaluronidase and collagenase. NE and cathepsin-G led to 3D-spheroid formation of ER+/E-CAD+ cells only. In 3D-spheroids from MCF7 cells grown with NE the luminal Epithelial Membrane Antigen (EMA) lined the external border of cell clusters, which faced cancer associated fibroblasts in co-cultures experiments, thus recapitulating the inverted polarity of MPCs. MCF7 3D-spheroids were tamoxifen resistant. Injection of NE in tumors of MCF7 cells in SCID mice triggered neoplastic cluster detachment, micropapillae, vascular emboli similar to 3D-spheorids and metastases. In a cohort of human breast carcinomas with LVI the MCF7-3D-spheroid-alike pattern was the most prevalent in tumor emboli. Immunohistochemical NE expression was mainly detected in polymorphous neutrophilic granulocytes (PMNs) within vessels and in the stroma and PMNs were significantly higher in breast carcinomas with LVI. In fully developed metastases within lymph-nodes, which reverted the EMA expression as in the primary tumor, no NE+ PMNs were observed. Our results may explain why high levels of NE negatively act on patient prognosis by creating a favorable environment for breast cancer invasion and metastasis. Citation Format: Caterina Marchio, Laura Annaratone, Davide Balmativola, Maria Stella Scalzo, Stefania Bolla, Silvia Grasso, Isabella Russo, Federica Fusella, Luigia Macrì, Mara Brancaccio, Paola Defilippi, Anna Sapino. Neutrophil elastase modulates breast cancer progression by fostering collective cell detachment and tumor emboli dissemination [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P1-07-27.

Correlation between microRNA-155 and WEE1 expression and their role in colon cancer

Objective To investigate the role of microRNA(miR)-155 and WEE1 in the tumorigenesis of colon cancer by detecting their expression level in tumor and normal tissue. Methods The expression level of miR-155 and WEE1 from tumor and normal tissues of 187 patients were detected by real-time fluorescent quantitative polymerase chain reaction(FQ-PCR).Correlation between variables was analyzed by Kendall'tau method.Survival curves were plotted using Kaplan-Meier method with Log-rank test.Cyclooxygenase(COX) proportional hazard model was used to evaluate the independent prognostic factors. Results miR-155 expression negatively and statistically significantly correlates with WEE1 expression in colon cancer tissue(r=-0. 555, P< 0. 01). miR-155 and WEE1 expression level was closely related with differentiation, TNM stage and lymphovascular metastasis(P< 0. 01).miR-155 and WEE1 expression level, lymphovascular metastasis were independent prognostic factors in colon cancer patients.The median overall survival difference between patients with increased level of miR-155 expression and normal levels of miR-155 expression had statistical significance(23 months vs.85 months,P<0. 05).The median overall survival difference between patients with decreased level of WEE1 expression and normal levels of WEE1 expression had statistical significance(25 months vs.70 months, P< 0. 05). Conclusion miR-155 and WEE1 expression level were independent prognostic factors of colon cancer patients. Decreased miR-155 expression and increased WEE1 expression predict a good prognosis. Key words: MicroRNA-155; WEE1; Colon cancer; Real-time polymerase chain reaction

Different expression of miRNAs in early esophageal squamous cell carcinoma with differential prognosis

Patients at the same pathological stage of early esophageal squamous cell carcinoma (ESCC) have significantly different prognoses. The aim of this study was to explore the relationship between specific microRNAs (miRNAs) in two groups of early ESCC patients who had different prognoses at the pT1N0 stage, and to study the prognostic significance of different miRNAs in early ESCC metastasis. We tried to identify prognostic markers that may be helpful in the selection of appropriate treatment for patients with early ESCC. We used TaqMan Human miRNA Arrays to detect and analyze bioinformatically the expression profiles of miRNAs in two groups, and the reverse transcription polymerase chain reaction method to verify the differences in miRNA expression. The miRNA arrays revealed a total of 29 markedly downregulated miRNAs in the survival group compared with the deceased group. Such miRNAs were associated with lympho-vascular invasion and metastasis, and acted as predictive markers of lympho-vascular invasion and metastasis. The detection of these miRNAs forms an important basis for the treatment of early ESCC. It can also help determine those patients with early ESCC who are good candidates for endoscopic resection treatment and those who need additional treatment after endoscopic resection.

Abstract 1159: Increased ceramide synthase 5 expression is associated with lymphovascular invasion, metastasis and poor survival in colorectal cancer.

Abstract Background: Ceramide synthase 5 (CerS5) is multi-pass transmembrane protein of the endoplasmic reticulum. Functioning as a bona fide (dihydro)ceramide synthase, CerS5 regulates the levels of short ceramide species. Ceramides are bioactive lipids implicated in proliferation, senescence, angiogenesis and death of normal and cancerous cells. We aimed to investigate CerS5 expression in colorectal cancer tissue and to evaluate the prognostic significance of CerS5 in colorectal cancer patients. Methods: CerS5 expression was analyzed by immunohistochemistry on tissue microarrays constructed from 125 colorectal cancer patients. The associations between CerS5 staining and the clinicopathological factors were examined using the Spearman rank correlation test. Patient survival was examined using the Kaplan-Meier survival analysis and Cox proportional hazard modeling, categorized according to high or low CerS5 staining intensity. Results: Immunohistological staining revealed an overexpression of membranous CerS5 in 63 out of 125 (50.4%) colorectal carcinomas. CerS5 overexpression significantly correlated with lymphovascular invasion and metastatic disease in matched tumor cases (P = 0.036 &amp; P = 0.037, respectively). Patients with CerS5 overexpressing tumors had a significantly lower overall 5-year survival (OS, P = 0.029) and 5-year recurrence-free survival (RFS, P = 0.047). Cox multivariate analysis showed that the increased CerS5 expression was an independent predictor of OS (hazard ratio (HR) = 1.604; 95% confidence interval (CI) 1.010-2.548, P = 0.045) and RFS (HR = 1.592; CI 1.036-2.447, P = 0.034). Conclusions: We found an increased expression of ceramide synthase 5 (CerS5) in 50% of investigated colorectal carcinomas, which was associated with lymphovascular invasion, metastasis and poor survival in colorectal cancer patients. Our study identifies CerS5 as a novel independent prognostic biomarker for patients with colorectal cancer. Citation Format: Seán Fitzgerald, Katherine M. Sheehan, Anthony O'Grady, Dermot Kenny, Richard O'Kennedy, Elaine W. Kay, Gregor Kijanka. Increased ceramide synthase 5 expression is associated with lymphovascular invasion, metastasis and poor survival in colorectal cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1159. doi:10.1158/1538-7445.AM2013-1159

32. Uncommon case of solid pseudopapillary tumour of the pancreas with secondary spread

Solid pseudopapillary tumor of the pancreas (SPT) is a rare neoplasm with low malignant potential and most commonly found in young women. With complete surgical excision of the tumor clinical outcome is usually favourable. We would like to present a very rare case of pseudopapillary tumour of the pancreas with extensive local and distant secondary spread. The patient underwent complex surgery and intraoperative chemotherapy. Increased malignancy of solid pseudopapillary tumours is usually associated with the following non-specific features: cellular polymorphism, elevated mitotic activity, peripancreatic tissue invasion, perineural and lymphovascular invasion and lymph node metastasis. The morphology of the presented tumor did not show most of the features described above except peripancreatic tissue invasion and distant spread.