Pancreatic melatonin enhances anti-tumor immunity in pancreatic adenocarcinoma through regulating tumor-associated neutrophils infiltration and NETosis

Tumor microenvironment contributes to poor prognosis of pancreatic adenocarcinoma (PAAD) patients. Proper regulation could improve survival. Melatonin is an endogenous hormone that delivers multiple bioactivities. Here we showed that pancreatic melatonin level is associated with patients' survival. In PAAD mice models, melatonin supplementation suppressed tumor growth, while blockade of melatonin pathway exacerbated tumor progression. This anti-tumor effect was independent of cytotoxicity but associated with tumor-associated neutrophils (TANs), and TANs depletion reversed effects of melatonin. Melatonin induced TANs infiltration and activation, therefore induced cell apoptosis of PAAD cells. Cytokine arrays revealed that melatonin had minimal impact on neutrophils but induced secretion of Cxcl2 from tumor cells. Knockdown of Cxcl2 in tumor cells abolished neutrophil migration and activation. Melatonin-induced neutrophils presented an N1-like anti-tumor phenotype, with increased neutrophil extracellular traps (NETs) causing tumor cell apoptosis through cell-to-cell contact. Proteomics analysis revealed that this reactive oxygen species (ROS)-mediated inhibition was fueled by fatty acid oxidation (FAO) in neutrophils, while FAO inhibitor abolished the anti-tumor effect. Analysis of PAAD patient specimens revealed that CXCL2 expression was associated with neutrophil infiltration. CXCL2, or TANs, combined with NET marker, can better predict patients' prognosis. Collectively, we discovered an anti-tumor mechanism of melatonin through recruiting N1-neutrophils and beneficial NET formation.

LncRNA LINC01857 drives pancreatic adenocarcinoma progression via modulating miR-19a-3p/SMOC2

Abstract Objectives Emerging evidence has demonstrated that LINC01857 exerts a pivotal function in many cancers. However, its function in Pancreatic Ductal Adenocarcinoma (PDAC) still remains unclear. This study was designed to investigate the regulatory character of LINC01857 in PDAC. Methods Bioinformatic tools and databases were used to seek potential miRNAs and mRNAs. Gene expression was evaluated by Reverse Transcription quantitative real-time Polymerase Chain Reaction (RT-qPCR), and western blot was used for protein level detection. A subcellular fraction assay was done to ascertain the location of LINC01857 in PANC-1 and BxPC-3 human pancreatic cancer cells. CCK-8, EdU, wound healing and Transwell assays were performed to inquire into the influence of LINC01857, and SPARC -related Modular Calcium-binding protein-2 (SMOC2) on cell viability, proliferation, migration, and invasion, respectively. The interaction between LINC01857 and its downstream genes was explored by RNA immunoprecipitation and luciferase reporter assays. Results LINC01857 levels were significantly elevated in PDAC. Knockdown of LINC01857 significantly restrained the proliferation, migration, invasion, and Epithelial-Mesenchymal Transition (EMT) process of PDAC cells. MiR-19a-3p was a downstream target of LINC01857, and miR-19a-3p levels were significantly decreased in PDAC cells. In addition, SMOC2 expression had a negative correlation with that of miR-19a-3p, and SMOC2 was a downstream target of miR-19a-3p. Furthermore, SMOC2 upregulation partially abolished the inhibitive influence of LINC01857 downregulation on cell proliferation, migration, invasion, and the EMT process. Conclusion LINC01857 promotes malignant phenotypes of PDAC cells via upregulation of SMOC2 by interacting with miR-19a-3p. HIGHLIGHTS LINC01857 is upregulated in PAAD and promotes malignant cellular behaviors. LINC01857 interacts with miR-19a-3p to regulate SMOC2 expression. LINC01857 promotes malignant cellular phenotypes by upregulating SMOC2.

Percutaneous Radiofrequency Ablation for Metachronous Hepatic Metastases after Curative Resection of Pancreatic Adenocarcinoma

OBJECTIVE: To retrospectively evaluate the safety and efficacy of percutaneous radiofrequency ablation (RFA) in patients with metachronous hepatic metastases arising from pancreatic adenocarcinoma who had previously received curative surgery.MATERIALS AND METHODS: Between 2002 and 2017, percutaneous RFA was performed on 94 metachronous hepatic metastases (median diameter, 1.5 cm) arising from pancreatic cancer in 60 patients (mean age, 60.5 years). Patients were included if they had fewer than five metastases, a maximum tumor diameter of ≤ 5 cm, and disease confined to the liver or stable extrahepatic disease. For comparisons during the same period, we included 66 patients who received chemotherapy only and met the same eligibility criteria described.RESULTS: Technical success was achieved in all hepatic metastasis without any procedure-related mortality. During follow-up, local tumor progression of treated lesions was observed in 38.3% of the tumors. Overall median survival and 3-year survival rates were 12 months and 0%, respectively from initial RFA, and 14.7 months and 2.1%, respectively from the first diagnosis of liver metastasis. Multivariate analysis showed that a large tumor diameter of > 1.5 cm, a late TNM stage (≥ IIB) before curative surgery, a time from surgery to recurrence of < 1 year, and the presence of extrahepatic metastasis, were all prognostic of reduced overall survival after RFA. Median overall (12 months vs. 9.1 months, p = 0.094) and progression-free survival (5 months vs. 3.3 months, p = 0.068) were higher in the RFA group than in the chemotherapy group with borderline statistical difference.CONCLUSION: RFA is safe and may offer successful local tumor control in patients with metachronous hepatic metastases arising from pancreatic adenocarcinoma. Patients with a small diameter tumor, early TNM stage before curative surgery, late hepatic recurrence, and liver-only metastasis benefit most from RFA treatment. RFA provided better survival outcomes than chemotherapy for this specific group with borderline statistical difference.

Clinical efficacy of DC-CIK immunotherapy in treatment of advanced pancreatic adenocarcinoma / 中国肿瘤生物治疗杂志

@#［ ［Abstract］ ］Objective: To analyze and compare the clinical efficacy and safety of dendritic cell cytokine-induced killer cells (DCCIK) combined with palliative therapy or chemotherapy in the treatment of advanced pancreatic carcinoma. Methods: A retrospective study was carried on 50 patients with advanced pancreatic carcinoma who were hospitalized in department of oncology of Shanxi Dayi Hospital during September 2012 to February 2016. The patients were divided into four groups according to the therapy they received (palliative treatment group, palliative+DC-CIK treatment group, chemotherapy group and chemotherapy+DC-CIK treatment group); the immunological function, quality of life and survival time of patients were analyzed; and the efficacy and safety of DC-CIK cell therapy was also evaluated. Results: The percentages of CD8+ T cells and NKT cells in DC-CIK combined therapy groups were significantly improved compared with that of pre-treatment, and the percentages of CD3+, CD8+, NK, NKT cells were increased compared with control groups (P<0.05). The quality of life of patients was significantly improved (P<0.05), while median PFS and median OS were improved but without statistical significance (P>0.05). Conclusion: Compared with palliative therapy and chemotherapy alone, combined DC-CIK immunotherapy can effectively improve the cellular immunity function and quality of life in patients with advanced pancreatic cancer. However, there was no significant extension in overall survival.

A New Modified FOLFIRINOX(mFOLFIRINOX)Regimen as First-Line Chemotherapy for Patient with Metastatic Pancreatic Adenocarcinoma(MPC) / 中山大学学报(医学科学版)

[Objective]To investigate efficacy and toxicity of a new modified FOLFIRINOX regimen(mFOLFIRI-NOX)as first-line chemotherapy for the patients with metastatic pancreatic adenocarcinoma(MPC).[Methods]20 patients with metastatic pancreatic adenocarcinoma(MPC)accepted mFOLFIRINOX arm(oxaliplatin 60 mg/m2,irinotecan 150 mg/m2,bolus of 5-FU 400 mg/m2,continuous infusion of 5-FU 2 400 mg/m2)first-line treatment and evaluated the curative effect and side effect after 3 cycles by imaging and laboratory at Sun Yat-sen Memorial Hospital from December 2012 to December 2016.The primary endpoint was overall survival(OS).The second endpoint was response rate(ORR) and toxicity.[Results]Four patients were excluded according to exclusion criteria. A total of 16 patients were enrolled including 7 males and 9 females.The median age is 55 years(ranging from 43 to 67 years).The median cycle of chemo-therapy was 4.5(3-15). The ORR was 31.3% including 5 patients with partial response,9 patients with stable disease and 2 patients with progression disease.The median follow-up duration was 14.1(2.7-24.5)months.The median OS was 16.6 months(95%CI,11.36-21.84). Overall survival rates at 6,12months were 80%and 55.6%,respectively. Three patients were presented grade 3-4 adverse events.[Conclusion]The new mFOLFIRINOX regimen improved survival of patients with MPC with tolerated toxicity as first line treatment.

Silencing pancreatic adenocarcinoma up-regulated factor increases the sensitivity of pancreatic cancer cell line to gemcitabine / 中华肝胆外科杂志

Objective To observe the influence on the sensitivity of pancreatic cancer cell line BxPC-3 to gemcitabine of silencing PAUF gene.Methods BxPC-3 cells,which overexpress PAUF,was stably transfected with PAUF-shCtrl and PAUF-shRNA to establish BxPC-3_shCtrl and BxPC-3_shPAUF cells as control and experiment group.Then the mRNA and protein expression level of PAUF in these two cell lines were detected by RT-PCR and western blot,respectively.The growth inhibition rates of these two cell lines treated with different concentrations of gemcitabine (0,3.1,6.25,12.5,25,50,100,200 nmol/L) were detected by MTT.Apoptosis rates in the cells treated with different concentrations of gemcitabine (0,75,100 nmol/L) were then observed by flow cytometry.Results The relative PAUF mRNA expression level in BxPC-3_shCtrl and BxPC-3 cells were 1.00 ± 0.06 and 0.83 ± 0.07,which were significantly high er than that in BxPC-3_shPAUF cells (0.25 ± 0.02;both P ＜ 0.05).The relative PAUF protein expression level in BxPC-3_shCtrl and BxPC-3 cells were 0.89 ± 0.07 and 0.95 ± 0.04,which were significantly high er than that in BxPC-3_shPAUF cells (0.31 ± 0.03;both P ＜ 0.05).The IC50 value of gemcitabine to BxPC-3_shCtrl cell was (22.88 ± 2.43) nmol/L,which was significantly higher than that of BxPC-3_shPAUF cells [(1.06 ± 0.02) nmol/L;P ＜ 0.05];apoptosis rate of BxPC-3_shPAUF cells treated by gemcitabine increased faster than that of BxPC-3_shCtrl cells.Conclusion PAUF silencing could greatly enhance the sensitivity of BxPC-3 cells to gemcitabine.

Analysis on Survival and Prognostic Factors in Patients with Resectable Pancreatic Adenocarcinoma / 华中科技大学学报（医学）（英德文版）

Survival after pancreatic cancer surgery is extremely unfavorable even after curative resection.Prognostic factors have been explored but remain largely undefined.The present study was to identify the role of clinical and laboratory variables in the prognostic significance of resectable pancreatic adenocarcinoma.A total of 96 patients who underwent curative resection for pancreatic cancer were included.Survival was evaluated based on complete follow-up visits and was associated with potential prognostic factors using the Kaplan-Meier method and Cox proportional hazard model survival analyses.The results showed that prognostic variables significantly reduced survival,including old age,poorly differentiated tumors,elevated tumor markers and positive lymph node metastasis (LNM).Age of older than 60 years (HR=l.83,P=0.04),LNM (HR=2.22,P=0.01),lymph node ratio (0＜LNR≤0.2,HR=1.38,P=0.042;LNR＞0.2,HR=1.92,P=0.017),initial CA199 (HR=4.80,P=0.004),and CEA level (HR=2.59,P=0.019) were identified as independent prognostic factors by multivariate analysis.It was concluded that LNR may be potent predictor of survival and suggests that surgeons and the pathologists should thoroughly assess lymph nodes prior to surgery.

Application of irreversible electroporation in the management of locally advanced pancreatic adenocarcinoma / 中华肝胆外科杂志

Irreversible electroporation (IRE) is a nontemperature based novel ablation therapy in which short highvoltage pulses are applied to treat tissues to permeabilize the cell membrane irreversibly and thus lead to cell apoptosis.Because it is characterized as non-thermal,IRE can reduce the risk of thermal damage to vital organs and structures so that it is more feasible for the treatment of unresectable pancreatic cancer and thus provide an ideal alternative to other thermal treatment modalities.This review aims to analyze current literature of IRE and to discuss the new progress of the application of IRE in the management of locally advanced pancreatic adenocarcinoma.

Inflammation and Cancer Development in Pancreatic and Biliary Tract Cancer / 대한소화기학회지

Chronic inflammation has been known to be a risk for many kinds of cancers, including pancreatic and biliary tract cancer. Recently, inflammatory process has emerged as a key mediator of cancer development and progression. Many efforts with experimental results have been given to identify the underlying mechanisms that contribute to inflammation-induced tumorigenesis. Diverse inflammatory pathways have been investigated and inhibitors for inflammation-related signaling pathways have been developed for cancer treatment. This review will summarize recent outcomes about this distinctive process in pancreatic and biliary tract cancer. Taking this evidence into consideration, modulation of inflammatory process will provide useful options for pancreatic and biliary tract cancer treatment.

A Case of Agenesis of the Dorsal Pancreas with Concomitant Pancreatic Ductal Adenocarcinoma in an Adult Patient / 대한췌담도학회지

Agenesis of dorsal pancreas (ADP) is an extremely rare congenital anomaly defined as an absence of the dorsal ductal system resulting from failure in the embryologic development of the pancreatic dorsal bud. Most of ADP patients are asymptomatic but some of them suffer recurrent pancreatitis and diabetes. Few number of pancreatic adenocarcinoma in association with ADP has been published previously in other countries. There was no such case reported in Korea. We report a case diagnosed as pancreatic adenocarcinoma with ADP.

Mechamism of PAUF in pancreatic cancer pathogenesis and progression / 中华肝胆外科杂志

Pancreatic adenocarcinoma up-regulated factor(PAUF),a newly discovered gene,is highly expressed in pancreatic cancer.PAUF promotes the metastasis and progression of pancreatic cancer through many ways,such as the activation of signal pathway (CXCR4,β-catenin,TPL2/MEK/ERK,FAK/Scr),increasing the adhesiveness of pancreatic cancer cells,promoting angiogenesis and vascular permeability.Simultaneously,CXCR4,β-catenin,TPL2/MEK/ERK and FAK/Scr are closely related with gemcitabine-resistance.Based on this theory,we infer that PAUF plays a role in gemcitabine-resistance of pancreatic cancer cells.So far,no related research has been done domestic and overseas.The research may find a clue for the mechanism of chemotherapy-resistance and provide a new target spot for the therapy of pancreatic cancer.

RE: Atypical Pulmonary Metastases from Pancreatic Adenocarcinoma

No abstract available.

Is a Preoperative Assessment of the Early Recurrence of Pancreatic Cancer Possible after Complete Surgical Resection?

BACKGROUND/AIMS: The prognosis of pancreatic adenocarcinoma (PAC) is poor. The serum carbohydrate antigen 19-9 (CA 19-9) level has been identified as a prognostic indicator of recurrence and reduced overall survival. The aim of this study was to identify preoperative prognostic factors and to create a prognostic model able to assess the early recurrence risk for patients with resectable PAC. METHODS: A series of 177 patients with PAC treated surgically at the St. Andrea Hospital of Rome between January 2003 and December 2011 were reviewed retrospectively. Univariate and multivariate analyses were utilized to identify preoperative prognostic indicators. RESULTS: A preoperative CA 19-9 level >228 U/mL, tumor size >3.1 cm, and the presence of pathological preoperative lymph nodes statistically correlated with early recurrence. Together, these three factors predicted the possibility of an early recurrence with 90.4% accuracy. The combination of these three preoperative conditions was identified as an independent parameter for early recurrence based on multivariate analysis (p=0.0314; hazard ratio, 3.9811; 95% confidence interval, 1.1745 to 15.3245). CONCLUSIONS: PAC patient candidates for surgical resection should undergo an assessment of early recurrence risk to avoid unnecessary and ineffective resection and to identify patients for whom palliative or alternative treatment may be the treatment of choice.

1-MT Enhances Potency of Tumor Cell Lysate-pulsed Dendritic Cells against Pancreatic Adenocarcinoma by Downregulating the Percentage of Tregs / 华中科技大学学报（医学）（英德文版）

This study examined whether 1-methyl-tryptophan[1-MT,an indoleamine 2,3-dioxygenase(IDO)inhibitor]could reduce CD4+CD25+regulatory T cells(Tregs)proliferation and improve the anti-tumor efficacy of dendritic cells(DCs)pulsed with tumor cell lysate in the mice bearing pancreatic adenocarcinoma.The models of pancreatic adenocarcinoma were established in C57BL/6 mice by subcutaneous injection of Pan02 cells.Eight mice which were subcutaneously injected with PBS served as control.The expression of IDO was determined in tumor draining lymph nodes(TDLNs)and spleens of the murine pancreatic adenocarcinoma models.The prevalence of Tregs was measured in the TDLNs and spleens before and after 1-MT administration.The dendritic cells were pulsed with tumor cell lysate for preparing DC vaccine.The DC vaccine,as a single agent or in combination with 1-MT,was administered to pancreatic adenocarcinoma mice.The anti-tumor efficacy was determined after different treatments by regular observation of tumor size.The results showed that the levels of IDO mRNA and protein in tumor-bearing mice were significantly higher than those in the normal control mice.The percentage of Tregs in the spleen and TDLNs was also higer in tumor-bearing mice than in normal control mice(P<0.05).Foxp3 expression was significantly lower in the TDLNs and spleens of tumor-bearing mice administrated with 1-MT than that in normal control mice.Furthemore,in the mice that were administered 1-MT plus DC vaccine,the tumor was increased more slowly than in mice treated with DC vaccine or 1-MT alone,or PBS on day 36(P<0.01).Our results indicated that 1-MT may enhance anti-tumor efficacy of dendritic cells pulsed with tumor cell lysate by downregulating the percentage of Tregs.

Transcatheter Arterial Chemoembolization for Hepatic Recurrence after Curative Resection of Pancreatic Adenocarcinoma

BACKGROUND/AIMS: Despite curative resection, hepatic recurrences cause a significant reduction in survival in patients with primary pancreatic adenocarcinoma. Transcatheter arterial chemoembolization (TACE) has recently been used successfully to treat primary and secondary hepatic malignancy. METHODS: Between 2003 and 2008, 15 patients underwent TACE because of hepatic recurrence after curative resection of a pancreatic adenocarcinoma. The tumor response was evaluated based on computed tomography scans after TACE. The overall duration of patient survival was measured. RESULTS: After TACE, a radiographically evident response occurred in six patients whose tumors demonstrated a tumor blush on angiography. Four patients demonstrated stabilization of a hypovascular mass. The remaining five patients demonstrated continued progression of hypovascular hepatic lesions. The median survival periods from the time of diagnosis and from the time of initial TACE were 9.6 and 7.5 months, respectively. CONCLUSIONS: TACE may represent a viable therapeutic modality in patients with hepatic recurrence after curative resection of pancreatic adenocarcinoma.

Effect of HIF-1α on VEGF-C Induced Lymphangiogenesis and Lymph Nodes Metastases of Pancreatic Cancer / 华中科技大学学报（医学）（英德文版）

The effect of hypoxia inducible factor-1 α (HIF-1α) on vascular endothelial growth factor C (VEGF-C) and the correlation between HIF-1α and lymphangiogenesis and lymph nodes metastases (LNM) in pancreatic cancer were investigated. Immunohistochemical SP method was used to detect the protein expression of HIF-1α and VEGF-C, and Lymphatic vessel density (LVD) was determined by stain of VEGFR-3, collagen type Ⅳ in 75 pancreatic head cancers from regional pancreatectomy (RP) during Dec. 2001 to Dec. 2003. The relationship between HIF-1α and VEGF-C, lymphangiogenesis, LNM was analyzed statistically. The results showed that the positive expression rate of HIF-1α and VEGF-C in pancreatic cancer tissues was 48.00 % (36/75) and 65.33 % (49/75) respectively. In positive group of HIF-1α, the positive rate of VEGF-C and LVD, and LVD rate was 80.56 % (29/36), 13.22±3.76 and 88.89 % (32/36) respectively, and in negative group of HIF-1α,positive rate of VEGF-C and LVD was 51.28 % (20/39), 5.98±2.17 and 66.67 % (26/39) respectively (P＜0.01 or P＜0.05). It was suggested that HIF-1α could promote the expression of VEGF-C, lymphangiogenesis and LNM in pancreatic cancer.

Prognostic Factors and Survival Rate of Pancreatic Adenocarcinoma after Curative Surgery

PURPOSE: This retrospective study aimed to identify the outcomes of resected pancreatic cancers and determine the prognostic factors for long term survival based on a single hospital experience. METHODS: Between January 1990 and February 2004, patients with a pancreatic ductal adenocarcinoma who had undergone resection at Severance hospital, Yonsei University, were analyzed retrospectively. RESULTS: Ninety-five patients underwent resection with curative intents for pancreatic adenocarcinoma. Sixty-seven (70.5%) patients had pancreatic head cancer, 27 (28.4%) cancers in the body and tail of the pancreas, and 1 had a diffusely spread type of pancreatic cancer. Procedures employed include Whipple resection (35.8%), pylorus preserving pancreaticoduodenectomy (33.7%), distal pancreatectomy (28.4%), and total pancreatectomy (2.1%). Stage Ia, Ib, IIa, IIb and III were present in 3.2, 4.2, 45.3, 44.2, and 3.2%, respectiely. The overall 5-year survival rate was 20.7%. Only low tumor stage was a significant predictive predictor of survival in univariate analysis (P<0.05). CONCLUSION: Long-term survival in patients with pancreatic adenocarcinoma is determined by the disease stage. This result suggests that early detection would be necessary to improve the survival of patients with pancreatic adenocarcinoma.

Ability of diagnosis of pancreatic adenocarcinoma by two dimensional ultrasound in comparing with CT scanner and endoscopic ultrasound

Studying on 35 patients were preoperative diagnosed as pancreatic adenocarcinoma received operations in ViÖt §øc hospital during 2000-2001 showed that: 29 patients with pancreatic adenocarcinoma and 6 patients without the disease. The two dimensional ultrasound found 18 cases (62%) with specificity (50%) and correct (60%), value of positive predict (85.7%). The rate of precise diagnosis of two dimensional ultrasound, CT scanner, and endoscopic ultrasound were 60%, 72.7%, and 83.3%, respectively.

The effect of fragile histidine triad gene on the growth and apoptosis of pancreatic adenocarcinoma cells / 中华消化杂志

Objective To study the effect of fragile histidine triad (FHIT) gene on pancreatic adenocarcinoma cells growth and tumorigenicity and explore the mechanism of FHIT gene in suppressing the deve lopment of pancreatic adenocarcinoma. Methods By the method of liposome transfection, pRC/CMV FHIT plasmid was transfected into 1990 cell lines which lose all of FHIT gene. Integration and expression of exogenous FHIT gene were confirmed by RT PCR and Western blot technique. 1990 pFHIT cell growth was observed in regular culture medium and tumorigenicity in nude mice. Its DNA was analyzed by electrophoresis. Results The growth of the cells transfected with FHIT gene (named as 1990 pFHIT cells) was suppressed significantly, and the tumorigenicity of the 1990 pFHIT cells was dramatically inhibited in nude mice as compared with that of the parental 1990 cells. Significantly increased apoptosis in 1990 pFHIT was found. Conclusions The growth and tumorigenicity of pancreatic adenocarcinoma cell can be inhibited by transduced exogenous FHIT gene. It's spectulated that FHIT suppress the development of pancreatic adenocarcinoma by the path of apoptosis.

Significance of Tumor Angiogenesis and p53 Protein Expression in Pancreatic Adenocarcinoma

PURPOSE: This study was designed to examine the microvessel count and p53 protein expression level in pancreatic adenocarcinoma patients and to determine how they correlate with the clinicopathologic factors and prognosis. In addition, the relationship between angiogenesis and p53 protein expression in pancreatic adenocarcinoma patients was investigated. METHODS: Paraffin-embedded tumor tissues from 30 patients with pancreatic adenocarcinomas that were completely removed by a radical resection were retrieved and analyzed. The vessels were immunostained with anti-factor VIII polyclonal antibodies, and 5 areas with the most discrete microvessels were counted under a 200 field. The mean was calculated, which was defined as the angiogenesis score (AS). p53 protein expression was detected by the immunohistochemical stain method. RESULTS: The mean AS was 37.7 13.6. There were no significant differences in sex, age, tumor size, histologic grade, T category (depth of invasion), N category (lymph node metastasis), pathologic stage and recurrence between the high and low AS group. The p53 protein expression rate was 73.3%, which did not correlate with the clinicopathologic factors and AS. However, recurrence was significantly related to the histologic grade and pathologic stage (p=0.040 and p=0.029). Among the various clinicopathologic factors, the N category (lymph node metastasis), the AS and recurrence (p=0.025, p=0.029 and p=0.003) were statistically significant according to a univariate survival analysis. In theCox regression analysis, the N category (lymph node metastasis) and recurrence were independent prognostic factors (odds ratio=4.344, 4.263). CONCLUSION: Although the survival rate in the high AS group was significantly lower than that in low AS group (p=0.029), the angiogenesis assessed by the microvessel count using immunohistochemical staining was not an independent prognostic factor in the Cox regression analysis.