Osteopontin influences the invasiveness of pancreatic cancer cells and is increased in neoplastic and inflammatory conditions

Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive malignancies, with an overall 5-year survival rate of less than 5%. Invasive tumor growth and early metastasis are two important reasons for this dismal prognosis. Osteopontin (OPN) is a secretory protein with a variety of functions, for example in cell adhesion and migration, inflammatory reaction and apoptosis. In this study the functional role of OPN in human pancreatic cancer and its potential use as a disease marker were analyzed. By real time quantitative PCR, there was a 2.2-fold and 1.6-fold increase of OPN mRNA in pancreatic cancers (n = 23) and chronic pancreatitis samples (n = 22), respectively, compared to normal pancreatic tissues (n = 20). Immunohistochemical analysis demonstrated OPN staining in 60% of the primary pancreatic tumors and in 72% of the lymph node and liver metastases. ELISA analysis of serum samples obtained from pancreatic cancer patients (n = 70), chronic pancreatitis patients (n = 12), and healthy donors (n = 20) showed a 1.6-fold increase in OPN serum levels in patients with tumors and a 1.9-fold increase in patients with chronic pancreatitis. Recombinant human OPN significantly increased the invasiveness of pancreatic cancer cells, without having any impact on cell proliferation. In addition, down regulation of OPN by specific siRNA molecules decreased pancreatic cancer cell invasion. In conclusion, OPN serum levels in pancreatic cancer and chronic pancreatitis patients are not significantly different, thereby restricting its role as a prognostic or follow-up marker. Our results do suggest, however, that blockade of OPN might be useful as a therapeutic approach to inhibit invasion and metastasis of pancreatic cancer cells.     

Down-regulation of BRCA1 in chronic pancreatitis and sporadic pancreatic adenocarcinoma.

PURPOSE: BRCA1 and BRCA2 are considered to be breast cancer susceptibility genes that may also contribute to pancreatic cancer development because family studies revealed mutation carriers to have an increased risk of developing pancreatic cancer. However, as demonstrated for breast and ovarian cancer, inactivation of BRCA in sporadic diseases is based on alteration in gene expression or functional alteration. EXPERIMENTAL DESIGN: To study a potential correlation of BRCA1 and BRCA2 to chronic pancreatitis and development of sporadic pancreatic adenocarcinoma, we have analyzed the expression of these genes by quantitative PCR and performed immunohistochemical analyses in normal pancreatic tissues, chronic pancreatitis, and pancreatic cancer specimens. RESULTS: BRCA1 expression was down-regulated in chronic alcoholic pancreatitis, in particular on the RNA level. Furthermore, our data indicate suppressed BRCA1 expression in pancreatic cancer on both the RNA and protein levels. Quantitative analysis of BRCA1 protein expression demonstrated regular staining in 50% of tumor specimens tested and reduced staining in 50% of tumor specimens tested. Correlation with the clinical outcome revealed a significantly better 1-year overall survival for patients with BRCA1-regular as compared with BRCA1-reduced or BRCA1-absent tumors. In contrast, no substantial differences in BRCA2 expression were found in chronic pancreatitis and pancreatic cancer samples. CONCLUSIONS: Our data demonstrate alteration of BRCA1 expression in chronic pancreatitis and sporadic pancreatic adenocarcinoma. We, for the first time, provide evidence for a role of BRCA1 in pancreatic carcinogenesis of noninherited tumors and for clinical outcome.     

结直肠癌黏蛋白1的表达及其与临床特征的相关性

目的:分析黏蛋白1(MUCs1)在结直肠癌患者中的表达情况及其与临床特征的关系。方法:选取2015年6月至2016年5月间在我院就诊的结直肠癌患者82例作为研究对象,以所有患者手术后的癌组织、正常结直肠组织为研究材料,提取组织中的总RNA,反转录后使用荧光定量PCR检测MUCs1 mRNA的表达情况,并分析结直肠癌患者TNM分期、术后6个月、12个月复发、转移、死亡情况与患者癌组织及正常结直肠组织中MUCs1 mRNA表达水平的关系。结果:随着患者T分期、N分期的逐渐提高,癌组织中MUCs1 mRNA的表达水平也逐渐升高,T1-T4、N0-N3分期患者癌组织中MUCs1 mRNA的表达水平与正常组织比较差异均具有统计学意义（P＜0.05）。术后6个月及术后12个月的复发、转移、死亡患者癌组织中MUCs1 mRNA的表达水平与正常组织相比较差异均具有统计学意义（P＜0.05）。术后6个月及术后12个月复发、转移、死亡患者癌组织中MUCs1 mRNA的表达水平与未复发、未转移、未死亡的患者相比较差异均具有统计学意义（P＜0.05）。结论:MUCs1在结直肠癌患者中的表达情况与预后呈明显的正相关性。     

胰腺癌:PET诊断学

Important challenges for imaging of pancreatic cancer are the late presentation of the disease and the fact that therapeutic management is of limited success. Surgery continues to be the only treatment that offers potential cure. Therefore, defining whether the patient has an operable tumor remains the ultimate aim of imaging in pancreatic cancer. PET and PET/ CT with fluorodeoxyglucose （FDG） are of value in differential diagnosis between pancreatitis and carcinoma and for the detection of remote metastases, but relatively inefficient in the detection of nodal disease. The detection of recurrent disease is of little clinical consequence. FDG-PET may be considered as a prognostic marker for patient survival or therapy response, but evidence for these applications is lacking. Future applications will broaden the spectrum of tracers applied using molecules for the assessment of proliferation and detection of receptors.     

胰腺癌组织中表皮生长因子受体、基质金属蛋白酶9的表达及其意义

目的 探讨胰腺癌组织中表皮生长因子受体(EGFR)、基质金属蛋白酶9(MMP-9)的表达及其 与胰腺癌临床病理特征的关系。方法 应用免疫组织化学SP法检测44例胰腺癌、相应癌旁胰腺组织、 13例慢性胰腺炎和7例正常胰腺组织中EGFR、MMP-9的表达,并分析其与临床病理特征的相关性。 结果 正常胰腺组织中均无EGFR及MMP-9的表达。慢性胰腺炎组织中EGFR、MMP-9的阳性表达 率分别为23.1%(3/13)和15.4%(2/13)。胰腺癌组织中EGFR、MMP-9的阳性表达率分别为61.4%(27/44) 和54.5%(24/44);相应癌旁组织中的阳性表达率分别为34.1%(15/44)和29.5%(13/44)。胰腺癌组织中 的EGFR及MMP-9阳性表达率均显著高于相应癌旁组织(P<0.05);胰腺癌组织及癌旁组织中EGFR及 MMP-9的表达均显著高于慢性胰腺炎组织和正常胰腺组织(P均<0.05)。EGFR和MMP-9的表达与胰腺 癌临床分期、分化程度、血管侵犯及淋巴结转移相关。结论 胰腺癌组织中EGFR的表达与MMP-9表 达密切相关,EGFR的表达与MMP-9的表达水平可作为了解胰腺癌生物学行为和判断预后的指标。     

干扰素-α诱导蛋白27在胰腺癌组织中的表达及临床意义北大核心

目的:探讨干扰素-α诱导蛋白27(IFI27)在胰腺癌组织中的表达情况及其临床意义。方法:免疫组织化学法检测IFI27蛋白在胰腺癌、癌旁及慢性胰腺炎组织中的表达情况,分析阳性表达与胰腺癌患者相关临床病理参数和预后的关系。利用生物信息学方法基于TCGA数据库分析IFI27在胰腺癌组织中的表达差异和预后相关性。结果:IFI27在胰腺癌组织中表达的阳性率高于癌旁以及慢性胰腺炎组织(P<0.05),且与血管侵犯、淋巴结转移以及肿瘤分化程度相关。胰腺癌患者的总体生存期与肿瘤分化程度、血管侵犯、淋巴结转移以及IFI27阳性表达相关。多因素分析显示IFI27阳性表达是胰腺癌患者预后的独立危险因素。基于TCGA数据库分析得出IFI27在胰腺癌组织中的表达量高于正常组织(P<0.05),生存曲线提示IFI27低表达患者的预后优于高表达患者(P<0.05)。结论:IFI27高表达的胰腺癌患者预后差,IFI27阳性表达是胰腺癌患者的独立预后指标。     

Pancreatic cancer–Diagnostics: PET

Important challenges for imaging of pancreatic cancer are the late presentation of the disease and the fact that therapeutic management is of limited success. Surgery continues to be the only treatment that offers potential cure. Therefore, defining whether the patient has an operable tumor remains the ultimate aim of imaging in pancreatic cancer. PET and PET/ CT with fluorodeoxyglucose (FDG) are of value in differential diagnosis between pancreatitis and carcinoma and for the detection of remote metastases, but relatively inefficient in the detection of nodal disease. The detection of recurrent disease is of little clinical consequence. FDG-PET may be considered as a prognostic marker for patient survival or therapy response, but evidence for these applications is lacking. Future applications will broaden the spectrum of tracers applied using molecules for the assessment of proliferation and detection of receptors.     

survivin基因在胰腺癌组织中的表达及与HSP27表达的关系

目的:探讨胰腺癌组织中survivin基因的表达及其与热体克蛋白(HSP27)表达的关系。方法:用RT-PCR法检测62例胰腺癌、12例慢性胰腺炎及10例正常胰腺组织中survivinmRNA的表达,免疫组织化学法检测胰腺癌组织中HSP27的表达。结果:survivin在胰腺癌中的表达率为74.2%,而在慢性胰腺炎及正常胰腺组织中未见表达;survivin表达与胰腺癌的肿块大小、分化程度、临床分期及淋巴结转移无相关性;survivin与HSP27的表达有相关性。结论:survivin在胰腺癌中过度表达,提示其在胰腺癌的发生和发展中起重要作用;survivin和HSP27通过抗凋亡机制协同参与胰腺癌的发生。     

Application of glycoscience to the early detection of pancreatic cancer

The prognosis of pancreatic cancer is extremely poor compared to other cancers. One of the reasons for this is the difficulty of early diagnosis. Surveillance using cancer biomarkers and image diagnosis can enable early detection and has improved the prognosis of hepatocellular carcinoma in Japan. However, it is very difficult to detect pancreatic cancer at an early stage using cancer biomarkers and image diagnosis alone. Fucosylation is one of the most important types of glycosylation involved in cancer and inflammation. We have developed a novel glycocancer biomarker, fucosylated haptoglobin (Fuc‐Hpt), and have investigated its usefulness for the diagnosis of pancreatic cancer over approximately 10 years. Recently, we also found that most pancreatic tissues surrounding pancreatic cancer exhibit chronic pancreatitis with fibrosis and/or fatty degeneration. Certain forms of chronic pancreatitis might indicate high risk for the development of pancreatic cancer. In this review, we provide a historical summary of our research on Fuc‐Hpt as a cancer biomarker, and discuss a potential early detection system for pancreatic cancer.     

Tumor microenvironment and progression of pancreatic cancer

Pancreatic ductal adenocarcinoma is characterized by ? tumor desmoplasia ?, a remarkable increase in connective tissue that penetrates and envelopes the neoplasm. It is becoming clear that this desmoplastic microenvironment of pancreatic cancer--which is forming approximately eighty percent of the tumor mass--is not a passive scaffold for the tumor cells but an active player in carcinogenesis. Several chemotherapeutic agents and novel molecular targeted therapies against epithelial tumor cells--although showing antitumor activity in cell culture and mouse experiments--have failed to show significant effects in the clinic. Thus, targeting pancreatic tumor cells alone seems unlikely to improve the dismal prognosis of pancreatic cancer. It has recently been shown that the activated stroma of pancreatic cancer is an independent prognostic marker with an impact on patient survival as much as the lymph node status of the cancer. Several primarily benign conditions associated with expansion of stromal and inflammatory components, such as chronic pancreatitis or hereditary pancreatitis are believed to increase the risk of pancreatic cancer. Similar observations have been made in other cancer types such as chronic hepatitis-liver cancer, Barrett dyplasia-esophageal cancer, and inflammatory bowel disease-colon cancer. The common denominator of all these conditions is; chronic inflammation leads to increased incidence of cancer. In this review the impact of the activated stroma on pancreatic carcinogenesis is discussed.     

Concentration and localization of carbohydrate antigen 19-9 in tissues of pancreatic cancer

Carbohydrate antigen (CA) 19-9 concentration in the tissue-extracts of cancerous and noncancerous pancreatic tissues were determined by radioimmunoassay. Pancreatic cancer tissues revealed significantly elevated CA 19-9 concentrations, when compared with chronic pancreatitis, normal adult pancreas, or fetal pancreas tissues. Metastatic liver tumors from pancreatic cancer also showed extremely high CA 19-9 concentrations, and there was no significant correlation between tissue CA 19-9 concentration and serum CA 19-9 level in patients with pancreatic cancer. In addition, positive localization of CA 19-9 was clearly observed in cancer cells of pancreatic cancerous tissues by immunohistochemical study, confirming the remarkable increase of CA 19-9 concentration in tissues of pancreatic cancer, although CA 19-9 was also partially found in non-malignant pancreatic tissues. The results indicated that CA 19-9 would be produced in great quantities by cancer cells in tissues of pancreatic cancer, and thus could be a valuable tumor associated antigen suggesting its clinical use as a tumor marker for cancer of the pancreas.     

Discrimination of metabolic profiles of pancreatic cancer from chronic pancreatitis by high-resolution magic angle spinning 1H nuclear magnetic resonance and principal components analysis

The metabolic profiles of Sprague-Dawley rat pancreases were investigated by high-resolution magic angle spinning proton magnetic resonance spectroscopy ((1)H NMR) combined with principal components analysis (PCA) to discriminate pancreatic cancer from chronic pancreatitis. Intact pancreatic tissue samples were obtained from Sprague-Dawley rats with histologically proven pancreatic cancer (n = 5), chronic pancreatitis (n = 5), and two matched controls (n = 5 per group). Two (1)H NMR experiments, single-pulse and Carr-Purcell-Meiboom-Gill, were carried out separately. Increases in phosphocholine and glycerophosphocholine levels and decreases in leucine, isoleucine, valine, lactate and alanine levels were observed in chronic pancreatitis, whereas the opposite trends were observed in pancreatic cancer. Increasing taurine and decreasing betaine were found both in chronic pancreatitis and in pancreatic cancer. Additionally, the lipid content in pancreatic cancer was higher than that in chronic pancreatitis. PCA was carried out for the single-pulse and Carr-Purcell-Meiboom-Gill (1)H NMR spectra, respectively, to visualize separation among the samples and to extract characteristic metabolites of pancreatic cancer and chronic pancreatitis. Decreased phosphocholine and glycerophosphocholine were suggested as unique metabolite indicators of pancreatic cancer. Furthermore, even with the disturbance of various quantities of lipid contents pancreatic cancer and chronic pancreatitis could be differentiated well by the combination of high-resolution magic angle spinning (1)H NMR and PCA. Thus this combination was demonstrated to have the potential to improve magnetic resonance spectroscopy for positive early diagnosis of pancreatic cancer in clinical settings.     

Expression of epidermal and transforming growth factors in pancreatic cancer

Pancreatic cancer continues to be a major clinical problem and little is known of the various cellular and molecular events associated with this malignancy. Growth factors and their receptors have important functions in the process of tumor progression. We have examined by immunocytochemistry, the expression of epidermal growth factor (EGF), its receptor (EGFR) and the transforming growth factors alpha and beta (TGF alpha and beta) in various grades of pancreatic adenocarcinoma. Expression of the growth factors was compared to their distribution in apparently normal pancreas and chronic pancreatitis. EGF, TGF alpha and TGF beta was expressed in normal pancreatic tissue while the expression of EGFR was slight and restricted. In chronic pancreatitis, this expression of EGFR increased and was found to be moderate in intensity. Expression of EGF, TGF alpha and TGF beta was similar to that seen in normal pancreas. Moderate to intense expression of EGF and TGF alpha was evident in all grades of pancreatic cancer. Expression of EGFR was intense in all these lesions. However, the most significant finding was the absence of TGF beta in all pancreatic cancer lesions. These results may have significant implications for pancreatic tumor progression. EGF and TGF alpha are growth promoters influencing the expression of EGFR. TGF beta, on the other hand exerts an anti-proliferative effect and favours differentiation. It therefore appears that the balance between EGF and TGF alpha on the one hand and TGF beta on the other may be critical in the process of tumor progression, especially if one considers chronic pancreatitis as a pre-malignant condition and the growth factor expression associated with it.     

Expression and potential function of the CXC chemokine CXCL16 in pancreatic ductal adenocarcinoma

CXC chemokines have a major influence on the angiogenesis, growth and metastatic potential of pancreatic ductal adenocarcinoma. CXCL16 is a unique transmembrane CXC chemokine, which is shed by members of the disintegrins and metalloproteases (ADAMs), in particular by ADAM10 and ADAM17. In our study, we evaluated expression and potential function of CXCL16 and its receptor CXCR6. CXCL16 and the receptor CXCR6 are upregulated in pancreatic ductal adenocarcinoma (PDAC) and chronic pancreatitis tissues in contrast to normal pancreatic tissues at the mRNA and protein levels. In 85 and 100% of the investigated samples, tumor cells showed positive immuno-staining for CXCL16 and CXCR6, respectively; furthermore, tubular complexes of chronic pancreatitis and the invasive front of PDAC were immunopositive for CXCL16 and CXCR6. Stimulation of PDAC cells with proinflammatory cytokines increased CXCL16 protein levels, whereas silencing of ADAM10 with siRNA transfection led to a decrease in CXCL16 protein levels in cell culture supernatants. No effects on cell viability were notable after incubation of cancer cells with CXCL16. However, CXCL16 markedly increased invasiveness of PDAC cells. Clinically, 82.5% of PDAC patients had higher CXCL16 serum values than the highest value seen in healthy donors. SELDI-TOF-MS analysis confirmed the upregulation of CXCL16 in sera of PDAC patients. In conclusion, CXCL16 in both transmembrane and soluble forms, and its receptor CXCR6, seem to play an important role in the pathobiology of pancreatic cancer and might be potential markers for pancreatic cancer diagnosis and a target for multimodal therapy concepts in the future.     

hnRNPF在胰腺癌中的表达及临床意义

目的:研究核内不均一性核糖核蛋白F(heterogeneous nuclear ribonucleoprotein F,hnRNPF)mRNA水平在胰腺癌中的表达及临床意义。方法:采用TCGA全基因组mRNA测序资料研究hnRNPF在消化系统肿瘤与正常组织间的差异,分析其表达水平与临床病理资料的相关性,并进行生存分析判断其预后价值。结果:hnRNPF在不同消化系统肿瘤中的表达较正常组织明显增高,差异具有统计学意义（P<0.001）,其中胰腺癌中差异倍数最大。hnRNPF在胰腺癌组织中的表达与肿瘤TNM分期(P=4.95E-03)、浸润深度(P=5.22E-03)相关,与年龄、性别、淋巴结浸润、慢性胰腺炎病史无统计学差异。hnRNPF在胰腺癌组织中的表达高低与总生存期有统计学意义（P=0.034 31）。结论:hnRNPF mRNA在不同的消化系统肿瘤中表达都增高,在胰腺癌中,hnRNPF高表达是预后不良的因素,可以作为判断预后的有效生物标志物。     

Cell lineage markers in human pancreatic cancer

The normal pancreas consists of three major cell types or lineages that share a common embryologic origin from pluripotent endodermal precursors. The type of cell that undergoes neoplastic transformation to form a pancreatic carcinoma is controversial and may influence the phenotype and biologic behavior of the tumor. In this study, immunohistologic techniques were used to determine the cell lineage differentiation expressed in 29 primary exocrine pancreatic adenocarcinomas, five metastatic exocrine pancreatic adenocarcinomas, and five islet cell neoplasma. Specimens of normal pancreas and chronic pancreatitis were used for comparison. The cell lineage markers consisted of monoclonal and polyclonal antibodies against trypsin and lipase (acinar cells); secretory component, carbonic anhydrase II, and pancreatic cancer mucin SPan-1 (ductal cells); and chromogranin-A and somatostatin (islet cells). The expression of carcinoembryonic antigen (CEA) and lysozyme were also determined. This collection of markers allowed the differentiation between acinar, ductal, and islet cells of normal pancreas and chronic pancreatitis specimens. The expression of cell lineage markers in islet cell tumors was homogeneous and restricted to chromogranin-A. In contrast, the expression of these markers in primary and metastatic exocrine pancreatic adenocarcinomas was variable. Reactivity with monoclonal anti-CEA was absent in normal pancreas, and was present in 83% of chronic pancreatitis specimens as well as 90% of exocrine pancreatic adenocarcinomas. In addition, lysozyme reactivity was absent in normal pancreas; however, lysozyme was expressed in one case of chronic pancreatitis, 17 cases of primary carcinoma, and three cases of metastatic carcinoma. These findings support the concept that the original transformed cell type in many pancreatic exocrine carcinomas resemble endodermal "stem cells" that retain the capability of differentiation along more than one cell lineage pathway.