Expression of p57/Kip2 protein in pancreatic adenocarcinoma

INTRODUCTION: Evaluation of the biologic character of carcinomas requires understanding of cell cycle regulators. AIMS: To investigate p57 expression in human pancreatic adenocarcinoma and cyst adenoma. METHODOLOGY: We examined the expression of p57(Kip2), a member of the Cip/Kip family, in 45 pancreatic adenocarcinomas, 7 cystadenomas, and 7 chronic pancreatitis cases. RESULTS: The p57 labeling index (LI) in duct epithelia in chronic pancreatitis averaged 32.8+/-8.3 and was significantly higher than in normal duct epithelia (18.8+/-6.6; p = 0.0011). For the carcinoma, the LI averaged 46.0+/-20.9, which was significantly higher than that for normal duct epithelia (p < 0.0001) and cystadenoma (16.0 11.2; p = 0.0007). However, it was significantly reduced in cases with stage IV disease (p = 0.0351), lymph node metastasis (p = 0.0003), larger size (p = 0.0094), capsular invasion (p = 0.0462), lymphatic invasion (p = 0.0351), and cell proliferating activity (p = 0.0002). In multivariate analysis, p57 LI in pancreatic adenocarcinoma was independently linked to high proliferating activity (p = 0.0230). CONCLUSION: These results suggest that p57 plays a role in the hyperplastic change of the ducts in chronic pancreatitis and that pS7 overexpression contributes to the downregulation of cell proliferation, and its decreased expression contributes to the progression of pancreatic adenocarcinoma.     

胰腺癌组织中Syk的表达及意义

目的探讨胰腺癌组织中脾酪氨酸激酶（Syk）的表达情况及意义。方法用免疫组化法和RT-PCR检测40例胰腺癌组织、9例慢性胰腺炎、8例正常胰腺组织中的Syk蛋白及其mRNA,形态计量学图象分析法测定微淋巴管密度（MLVD）。结果Syk蛋白在胰腺癌、慢性胰腺炎和正常胰腺组织中阳性率分别为27.5%、77.8%和100%;Syk蛋白与胰腺癌TNM分期、分化程度和淋巴结转移有关（P〈0.05）。Syk mRNA在胰腺癌中呈低表达（P〈0.01）。胰腺癌组织中微淋巴管密度（MLVD）在TNM分期III-IV期组显著高于I-II期组（P〈0.05）,淋巴结转移阳性组显著高于阴性组（P〈0.01）。Syk蛋白阴性表达者的MLVD显著高于阳性组（P〈0.01）。结论Syk在胰腺癌中表达减低或缺失,与胰腺癌的发展显著相关。     

甲基转移酶3B基因在胰腺癌中的表达

目的 检测胰腺癌组织中甲基转移酶3B(DNMT3B)基因表达,分析其与胰腺癌临床病理参数的关系.方法 应用实时定量PCR和免疫组织化学方法检测42例胰腺癌组织及相应癌旁组织、10例正常胰腺组织中DNMT3B mRNA和蛋白表达.结果 胰腺癌组织、癌旁组织和正常胰腺组织DNM33B mRNA表达量分别为9.4±5.9、1.02±0.71和0,相差非常显著(P<0.05);DNMT3B蛋白表达阳性率分别为83.3%、14.3%和10.0%,相差也非常显著(P<0.01).DNMT3B mRNA表达与I临床分期、肿瘤分化程度和淋巴结转移密切相关(P<0.05或P<0.01);DNMT3B蛋白表达与肿瘤的部位、淋巴结转移有关(P<0.01或P<0.05);DNMT3B mRNA和蛋白表达均与年龄、性别、神经浸润、肿瘤大小、血CEA和CA19-9浓度无关.结论 胰腺癌DNMT3B mRNA和蛋白高表达提示胰腺癌已发生淋巴结转移,预后较差.     

甲基转移酶3B基因在胰腺癌中的表达

目的检测胰腺癌组织中甲基转移酶3B（DNMT3B）基因表达,分析其与胰腺癌临床病理参数的关系。方法应用实时定量PCR和免疫组织化学方法检测42例胰腺癌组织及相应癌旁组织、10例正常胰腺组织中DNMT3B mRNA和蛋白表达。结果胰腺癌组织、癌旁组织和正常胰腺组织DNMT3B mRNA表达量分别为9．4±5．9、1．02±0．71和0,相差非常显著（P〈0．05）;DNMT3B蛋白表达阳性率分别为83．3％、14．3％和10．0％,相差也非常显著（P〈0．01）。DNMT3B mRNA表达与临床分期、肿瘤分化程度和淋巴结转移密切相关（P〈0．05或P〈0．01）;DNMT3B蛋白表达与肿瘤的部位、淋巴结转移有关（P〈0．01或P〈0．05）;DNMT3B mRNA和蛋白表达均与年龄、性别、神经浸润、肿瘤大小、血CEA和CA19—9浓度无关。结论胰腺癌DNMT3B mRNA和蛋白高表达提示胰腺癌已发生淋巴结转移,预后较差。     

胰腺癌组织表皮生长因子mRNA表达的意义

目的探讨ＥＧＦｍＲＮＡ的表达与胰腺癌发生、发展及预后的关系．方法应用Ｎｏｒｔｈｅｒｎｂｌｏｔ杂交方法，检测胰腺癌２７例和正常胰组织７例中ＥＧＦｍＲＮＡ表达．结果正常胰腺组织７例未检测到ＥＧＦｍＲＮＡ表达，胰腺癌２７例ＥＧＦｍＲＮＡ阳性表达率６６７％（１８例），经卡方检验发现，ＥＧＦｍＲＮＡ表达与胰腺癌病理分级、临床分期及伴随局部淋巴结转移呈显著相关（Ｐ＜００５）．结论ＥＧＦｍＲＮＡ表达与胰腺癌发生有关，胰腺癌ＥＧＦｍＲＮＡ表达可作为胰腺癌预后的参考指标     

胰腺癌中FXYD6蛋白的表达及临床意义

目的:探讨FXYD6蛋白在胰腺癌中的表达及临床意义.方法:应用免疫组化法检测FXYD6蛋白在胰腺癌中的表达,并分析其与临床病理因素的关系.结果:47例胰腺癌病例中,FXYD6蛋白阳性表达42例,阳性表达率为89.34%,而正常胰腺组织中不表达或低表达,两组相比有显著性差异(X2=10.863l,P=0.0125).按FXYD6蛋白表达程度将47例胰腺癌病例分为高表达组和低表达组,两组年龄、性别、肿瘤大小及远处转移之间的差异无统计学意义,而肿瘤分化程度(X=11.562,P=0.0031)、淋巴转移(X2=4.1947,P=0.0406)、TNM分期(X2=9.5995,P=0.0223).间的差异有显著统计学意义,肿瘤分化程度低、出现淋巴转移、TNM分期为Ⅲ.Ⅳ期的胰腺癌组织中FXYD6蛋白表达明显增高.Spearman等级相关分析显示,FXYD6蛋白表达程度与肿瘤大小(r=0.2991)、分化程度(r=0.6980)、淋巴转移(r=0.4971)、远处转移相关(r=0.4967).结论:FXYD6蛋白在胰腺癌发病机制中发挥重要作用.     

Expression of E-cadherin,alpha-and beta-catenins in patients with pancreatic adenocarcinoma

Background/Aims: E-Cadherin and its associated cytoplasmic proteins, catenins, are important mediators of epithelial cell-cell adhesion and intracellular signaling. Much evidence exists suggesting a tumor invasion suppressor role for E-cadherin and catenins and loss of expression, as well as mutations, has been described in a number of epithelial cancers. The aim of this study was to evaluate the expression of E-cadherin and catenins in pancreatic adenocarcinoma tissue, and to examine the relationship between these expression and various clinicopathological parameters. Methods: In this study, we conducted an immunohistochemical investigation of expression of E-cadherin, α- and β-catenins in 30 tissue samples obtained from pancreatic ductal adenocarcino-ma patients undergoing surgical treatment. Results: In the pancreatic mucosa of noncancerous areas, epithelial cells showed equally strong membranous expression of E-cadherin, α- and β-catenin proteins at the cell-cell boundaries. Reduced expression of E-cadherin, α- and β-catenins was demonstrated in 60.0, 40.0, and 56.7% of cancer tissues, respectively. Reduced expression of E-cadherin, α- and β-catenins correlated with tumor dedifferentiation (p = 0.012, 0.013, and 0.033, respectively). Reduced expression of E-cadherin correlated with stage and lymph node involvement (p = 0.031, 0.009, respec-tively). α-Catenin and β-catenin expression did not correlate with the patient's age and sex, with the tumor size, location, stage and depth of invasion, or lymph node involvement and distant metastasis. Conclusion: These results suggest that the E-cadherin and catenins may be a useful marker of differentiation and prognosis in pancreatic adenocarcinoma, although the mechanisms underlying changes in E-cadherin and catenin expression are not fully known.     

12-脂氧合酶在胰腺癌中的表达及其临床意义

目的 检测胰腺癌组织和胰腺癌细胞株SW1990和PANC1的12-脂氧合酶(12-lipooxygenase,12-LOX)表达,探讨其与胰腺癌临床病理参数的关系.方法 分别应用免疫组化染色、RT-PCR和蛋白质印迹法检测胰腺癌组织、癌旁正常胰腺组织及胰腺癌细胞株SW1990和PANC1中12-LOX mRNA和蛋白的表达.分析胰腺癌组织12-LOX表达与临床病理参数的相关性.结果 30例胰腺癌组织12-LOX表达阴性8例,弱阳性7例,强阳性15例,总阳性率为73.3％.SW1990和PANC1细胞均表达12-LOX.阳性表达的胰腺癌组织及两株胰腺癌细胞株均表达12-LOX mRNA,而癌旁正常胰腺组织不表达12-LOX mRNA及蛋白.胰腺癌组织12-LOX强阳性表达与肿瘤TNM分期、肿瘤病理分级和淋巴结转移相关(P＜0.05),而与患者年龄、性别无关.结论 12-LOX在胰腺癌中表达上调,与胰腺癌的恶性生物学行为有关.     

Expression of heparin-binding epidermal growth factor-like growth factor in pancreatic adenocarcinoma.

BACKGROUND: Previous studies demonstrated that heparin-binding epidermal growth factor-like growth factor (HB-EGF) contributes to carcinogenesis and carcinoma progression. In this study, we investigated its expression in human pancreatic adenocarcinoma. METHODS: We immunohistochemically investigated the expression of HB-EGF in 40 cases of pancreatic adenocarcinoma. RESULTS: HB-EGF was only occasionally and faintly expressed in normal and hyperplastic pancreas duct epithelia. In pancreatic adenocarcinoma, 22 (55.0%) of the 40 cases were classified as positive for HB-EGF. Its expression was more frequently observed in cases with a low Ki-67 labeling index, well differentiated. early stage, small size, without lymph node metastasis and low EGF-R expression. CONCLUSION: These results suggest that HB-EGF mainly plays a role in early phase of the progression of pancreatic adenocarcinoma.     

Expression of inducible nitric oxide synthase and cyclooxygenase-2 in pancreatic adenocarcinoma： Correlation with microvessel density

AIM: Cyclooxygenases (COX) are key enzymes for conversion of arachidonic acid to prostaglandins. Nitric oxide synthase (NOS) is the enzyme responsible for formation of nitric oxide. Both have constitutive and inducible isoforms. The inducible isoforms (iNOS and COX-2) are of great interest as regulators of tumor angiogenesis, tumorigenesis and inflammatory processes. This study was to clarify their role in pancreatic adenocarcinomas. METHODS: We investigated the immunohistochemical iNOS and COX-2 expression in 40 pancreatic ductal adenocarcinomas of different grade and stage. The results were compared with microvessel density and clinicopathological data. RESULTS: Twenty-one (52.5%) of the cases showed iNOS expression, 15 (37.5%) of the cases were positive for COX-2. The immunoreaction was heterogeneously distributed within the tumors. Staining intensity was different between the tumors. No correlation between iNOS and COX-2 expression was seen. There was no relationship with microvessel density. However, iNOS positive tumors developed more often distant metastases and the more malignant tumors showed a higher COX-2 expression. There was no correlation with other clinicopathological data. CONCLUSION: Approximately half of the cases expressed iNOS and COX-2. These two enzymes do not seem to be the key step in angiogenesis or carcinogenesis of pancreatic adenocarcinomas. Due to a low prevalence of COX-2 expression, chemoprevention of pancreatic carcinomas by COX-2 inhibitors can only achieve a limited success.     

The clinical utility of immunoglobulin G4 in the evaluation of autoimmune pancreatitis and pancreatic adenocarcinoma

Background

Elevation in the serum immunoglobulin-G4 (IgG4) level has been used as a diagnostic marker to distinguish autoimmune pancreatitis (AIP) from pancreatic ductal adenocarcinoma (PDAC), but its true utility is ill-defined. This study evaluates the clinical utility of IgG4 in differentiating AIP from PDAC.

Expression of tissue factor in pancreatic adenocarcinoma is associated with activation of coagulation

AIM: To study expression of tissue factor (TF) in pancreatic cancer and its role in the development of thromboembolism. METHODS: TF expression was studied in eight human pancreatic carcinoma cell lines by Northern blot and indirect immunofluorescence. Expression of alternatively spliced TF (asTF) was assessed by RT-PCR. In addition, TF expression was determined by immunofluorescence in pancreatic tissues of 19 patients with pancreatic adenocarcinoma (PCa), 9 patients with chronic pancreatitis (CP) and 20 normal controls. Plasma samples (30 PCa-patients, 13 CP-patients and 20 controls) were investigated for soluble TF levels and coagulation activation markers [thrombin-antithrombin III complex (TAT), prothrombin fragment 1 2 (F1 2)]. RESULTS: All pancreatic carcinoma cell lines expressed TF (8/8) and most of them expressed asTF (6/8). TF expression at the protein level did not correlate with the differentiation of the carcinoma cell line. All but two pancreatic cancer tissue samples stained positive for TF (17/19). In all samples of CP weak staining was restricted to pancreatic duct cells, whereas only a few subendothelial cells were positive in 9/20 of normal controls. TF and TAT levels in PCa patients were significantly elevated compared to controls whereas elevated F1 2 levels did not reach statistical significance compared to controls. In CP patients TAT and F1 2 levels proved to be significantly elevated compared to controls, although TAT elevation was less pronounced than in PCa patients. CONCLUSION: We conclude that in addition to the upregulated expression of TF on the cell membrane, soluble TF might contribute to activation of the coagulation system in pancreatic cancer.     

Expression of tissue factor in pancreatic adenocarcinoma is associated with activation of coagulation

AIM: To study expression of tissue factor (TF) in pancreatic cancer and its role in the development of thromboembolism. METHODS: TF expression was studied in eight human pancreatic carcinoma cell lines by Northern blot and indirect immunofluorescence. Expression of alternatively spliced TF (asTF) was assessed by RTPCR. In addition, TF expression was determined by immunofluorescence in pancreatic tissues of 19 patients with pancreatic adenocarcinoma (Pca), 9 patients with chronic pancreatitis (CP) and 20 normal controls. Plasma samples (30 Pca-patients, 13 CP-patients and 20 controls) were investigated for soluble TF levels and coagulation activation markers [thrombin-antithrombin Ⅲ complex (TAT), prothrombin fragment 1 + 2 (F1 + 2)]. RESULTS: All pancreatic carcinoma cell lines expressed TF (8/8) and most of them expressed asTF (6/8). TF expression at the protein level did not correlate with the differentiation of the carcinoma cell line. All but two pancreatic cancer tissue samples stained positive for TF (17/19). In all samples of CP weak staining was restricted to pancreatic duct cells, whereas only a few subendothelial cells were positive in 9/20 of normal controls. TF and TAT levels in Pca patients were significantly elevated compared to controls whereas elevated F1 + 2 levels did not reach statistical significance compared to controls. In CP patients TAT and F1 + 2 levels proved to be significantly elevated compared to controls, although TAT elevation was less pronounced than in Pca patients. CONCLUSION: We conclude that in addition to the upregulated expression of TF on the cell membrane, soluble TF might contribute to activation of the coagulation system in pancreatic cancer.     

Perspectives in the treatment of pancreatic adenocarcinoma

Pancreatic ductal adenocarcinoma(PDAC) is an incurable lethal disease whose incidence rate is growing. There is no effective screening for detection of early stage tumors and,in most cases,PDAC is diagnosed at advanced disease stages,when radical pancreatic resection is not possible. The aggressive nature of pancreatic tumor cells lies in the complex genetic mechanisms behind their uncontrolled capability to grow and metastasize,which involve essential adaptive changes in cellular metabolism,signaling,adhesion and immunoediting. In addition,PDAC cells promote a dense functional stroma that facilitates tumor resistance to chemotherapy and radiation. During the last two decades,gemcitabine has been the reference for the systemic treatment of PDAC. However,recently,a regimen combining fluorouracil,irinotecan,oxaliplatin,and leucovorin(FOLFIRINOX) and another combining albumin-bound paclitaxel with gemcitabine have shown clear therapeutic advantage in advanced PDAC,with survival outcomes of 11.3 and 8.5 mo on phase Ⅲ trials,respectively,over singleagent gemcitabine. With the pending issue of their higher toxicities,these regimens set the reference for ongoing and future clinical studies in advanced PDAC. In addition,the efficacy of oral fluoropyrimidine(S-1) has been well documented in Asiatic PDAC patients. The development of therapeutic approaches other than cytotoxic drugs has proven difficult in the past,with only one drug(erlotinib) approved to date. Besides,a number of agents targeting signaling pathways in tumor or stroma cells are being investigated. Likewise,immunotherapies that target PDAC in various ways are the subject of a number of clinical trials. The search for reliable biomarkers with diagnostic and prognostic value using genomics and mass spectrometry methods may facilitate monitoring and refinement of therapies. This review focuses on current understanding of the pathogenesis of PDAC and the latest developments in the treatment of advanced PDAC.     

Expression of fibroblast activation protein in human pancreatic adenocarcinoma and its clinicopathological significance

AIM: To examine fibroblast activation protein (FAP) expression in pancreatic ductal adenocarcinoma (PDAC) and to analyze its relationship with the clinicopathology of PDAC.METHODS: FAP expression was examined in 134 PDAC specimens by immunohistochemistry, and in four pancreatic cancer cell lines (SW1990, Miapaca-2, AsPC-1 and BxPC-3) by Western blotting assay. We also analyzed the association between FAP expression in PDAC cells and the clinicopathology of PDAC patients.RESULTS: The results showed that the FAP was ex-pressed in both stromal fibroblast cells (98/134, 73.1%) and carcinoma cells (102/134, 76.1%). All 4 pancreatic cancer cell lines expressed FAP protein at different levels. Protein bands corresponding to the proteolytically active 170-kDa seprase dimer and its 88-kDa seprase subunit were identified. Higher FAP expression in carcinoma cells was associated with tumor size (P < 0.001), fibrotic focus (P = 0.003), perineural invasion (P = 0.009) and worse clinical outcome (P = 0.0085).CONCLUSION: FAP is highly expressed in carcinoma cells and fibroblasts in PDAC tissues, and its expression is associated with desmoplasia and worse prognosis.     

TGF-β/Smads信号通路相关蛋白在胰腺上皮内瘤变和胰腺癌组织中的表达

目的探讨转化生长因子(TGF)-β/Smads信号转导通路中Smads相关蛋白、TGF-β1及其Ⅰ、Ⅱ型受体在胰腺上皮内瘤变(PanIN)和胰腺癌组织中表达的意义.方法用EnVision和SP免疫组化技术检测266灶不同级别PanINs和121例胰腺癌组织中Smads相关蛋白、TGF-β1及其Ⅰ、Ⅱ型受体的表达并联系临床病理学指标进行相关分析.结果高级别PanINs病灶Smad4表达率(60.6%,20/33)显著低于低级别PanINs Smad4表达率(79.8%,186/233)(P<0.05);而高级别PanINs中Smad7、TGF-β1、TGF-βRⅡ表达率明显高于低级别PanINs(P<0.05).胰腺癌组织中,Smad4蛋白表达率在淋巴结转移组和神经受累组显著低于各自对照组(P<0.05);Smad7、TGF-β1及其Ⅰ、Ⅱ型受体的表达率在淋巴结转移组和神经受累组则分别显著高于各自对照组(P<0.05).胰腺上皮内瘤变和胰腺癌组织中Smad2、4、7蛋白,TGF-β1及其Ⅰ、Ⅱ型受体表达率的差异具有非常显著性(P<0.01).结论从低级别PanINs到高级别PanINs再到胰腺癌中,Smad4蛋白表达率逐渐降低,而Smad7、TGF-β1及其Ⅰ、Ⅱ型受体表达率逐渐升高,支持胰腺癌形成的分子模型.     

Syk和VEGF-C在胰腺癌组织中的表达及临床意义

目的进一步探讨胰腺癌发生、发展机制。方法收集4JD例胰腺癌、9例慢性胰腺炎、8例正常胰腺组织标本,应用免疫组织化学方法和RT—PCR检测脾酪氨酸激酶（Syk）和血管内皮生长因子-C（VEGF—C）表达情况,分析两者表达与胰腺癌临床病理特征的相关性。结果Syk和VEGF—C在胰腺癌组织中的表达显著低于在正常胰腺组织中的表达（P〈0．01、0．05）,并与胰腺癌TNM分期、分化程度和淋巴结转移有关（P〈0．05）,两者表达具有显著相关性（P=0．019）。结论Syk、VEGF-C在胰腺癌发生、发展中可能具有协同作用。