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The Role of Notch3 and Related Pathways in Pancreatic Cancer

By Helena Doucas


Introduction: Pancreatic cancer is a malignancy with a very poor prognosis, and therefore insights into the derangement in cell signalling pathways which result in cancer development may provide important information in understanding the disease process, and in developing future therapeutic strategies. The aim of this study was to examine the expression and potential importance of several intracellular signalling proteins. Particular attention was given to Notch3.\ud Methods: An immunohistochemical study was performed on 23 samples of human pancreatic adenocarcinorna and 12 samples of normal pancreas, to assess expression of Notch3, cyclin D1, pAkt, STAT3 and pSTAT3, β-catenin, and pin1. The degree of staining, and intracellular locations of the proteins were assessed. Immunohistochemical data were correlated with clinical parameters, including tumour resectability, size, presence of metastases, and plasma levels of CA19-9. In vitro studies were also performed on two pancreatic cell lines. Basal levels of protein expression were assessed by western blotting. The effect of Notch inhibition by γ-secretase inhibitors and by transfecting cells with Notch3 siRNA was also examined. Cell survival studies were performed using the Annexin V apoptosis assay.\ud Results: Pin1 was not overexpressed in pancreatic adenocarcinoma. However, Notch3 was significantly overexpressed in the cytoplasm of 73.9% of tumours. Nuclear expression, never observed in normal ductal tissue, was noted in 43.5% of cases. No tumour expressing nuclear Notch3 was surgically resectable, and there was a significant correlation between nuclear Notch3 staining and increasing levels of CA19-9. There were also significant correlations between expression and intracellular location of Notch3 and each of STAT3, pSTAT3, and pAkt, but not cyclin D1. Notch3 was expressed in untreated ASPC1 cells, but not in PANC1 cells. Its levels could be reduced using either γ-secretase inhibitors or siRNA. Notch inhibition also resulted in a fall in levels of pAkt, but initial experiments did not demonstrate an effect upon cell survival.\ud Conclusion: The presence of Notch3 in tumour nuclei is likely to represent functional activation of the protein, and is clearly linked to more aggressive tumours. In the future, targeting the Notch pathway may prove to be of therapeutic benefit

Publisher: University of Leicester
Year: 2006
OAI identifier: oai:lra.le.ac.uk:2381/8469

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  1. (1996). A human peptidyl-prolyl isomerase essential for regulation of mitosis.
  2. (2002). A multi-institutional study of immunohistochernical investigation for the roles of cyclin D1 and E-cadherin in superficial squamous cell carcinoma of the esophagus.
  3. (1999). A presenilin-1 -dependent gamma-secretase-like protease mediates release of Notch intracellular domain.
  4. (2004). A signalling pathway controlling c-Myc degradation that impacts oncogenic transformation of human cells.
  5. (2003). Activated Notchl inhibits p53-induced apoptosis and sustains transformation by human papillornavirus type 16 E6 and E7 oncogenes through a P13K-PKB/Akt-dependent pathway.
  6. (2002). Activation of AKT/PKB in breast cancer predicts a worse outcome among endocrine treated patients.
  7. (2001). Activation of c-MYC and c-MYB protoChapter 6. References oncogenes is associated with decreased apoptosis in tumor colon progression. Anticancer Res,
  8. (2002). Activation of Notch-1 signaling maintains the neoplastic phenotype in human Ras-transformed cells. Nat Med,
  9. (2003). Adjuvant therapy in pancreatic cancer: historical and current perspectives.
  10. (2003). Advanced Pancreatic Ductal Cancer: Fibrotic Focus and beta-Catenin Expression Correlate With Outcome.
  11. (2003). AKT inhibition is associated with chemosensitisation in the pancreatic cancer cell line MIA-PaCa2.
  12. (1995). Alterations in Notch signaling in neoplastic lesions of the human cervix.
  13. (1998). Amplification and overexpression of cyclin D1 in aggressive human esophageal cancer. Oncol Rep,
  14. (1992). An immunochernical analysis of the human nuclear phosphoprotein p53. New monoclonal antibodies and epitope mapping using recombinant p53.
  15. (2002). Analysis of the beta-catenin/T cell factor signaling pathway in 36 gastrointestinal and liver cancer cells.
  16. (1991). Antigen retrieval in formalin-fixed, paraffinembedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections.
  17. (1995). Association of p53 gene mutations With short survival in pancreatic adenocarcinorna.
  18. (2003). Beta-catenin accumulation in the progression of human hepatocarcinogenesis correlates with loss of E-cadherin and accumulation of p53, but not With expression of conventional WNT-1 target genes.
  19. (2002). Beta-catenin and cyclin DI expression in human hepatocellular carcinoma. Oncol Rep,
  20. (2002). Beta-catenin expression and allelic loss at APC in sporadic colorectal carcinogenesis. Virchows Arch,
  21. (1999). Beta-catenin regulates expression of cyclin D1 in colon carcinoma cells.
  22. (2003). Blockade of constitutively activated Janus kinase/signal transducer and activator of transcription-3 pathway inhibits growth of human pancreatic cancer.
  23. (2002). c-MYC activation in primary and metastatic ductal adenocarcinoma of the pancreas: incidence, mechanisms, and clinical significance. Mod Pathol,
  24. (2001). C-myc amplification and cluster analysis in human gastric carcinoma. Anticancer Res,
  25. (1992). Carcinoma of the pancreas and papilla of Vater. presenting symptoms, signs, and diagnosis related to stage and turnour site. A prospective multicentre trial in 472 patients. Norwegian Pancreatic Cancer Trial.
  26. (2003). Catalytic site-directed gamma-secretase complex inhibitors do not discriminate phan-nacologically between Notch S3 and beta-APP cleavages.
  27. (2004). Chapter 6. References JAGGEDI expression is associated with prostate cancer metastasis and recurrence.
  28. (2000). Clinical significance of matrix metal loprotei nase-7 expression in esophageal carcinoma.
  29. (1993). Comparison of mutations of ras oncogene in human pancreatic exocrine and endocrine tumors.
  30. (2001). Copy number of cancer genes predict tumor grade and survival of pancreatic cancer patients. Anticancer Res,
  31. (2001). Cyclin DI expression in transitional cell carcinoma of the bladder: correlation with p53, wafl, pRb and Ki67.
  32. (2002). Cyclin DI overexpression in colorectal carcinoma in vivo is dependent on beta-catenin protein dysregulation, but not k-ras mutation.
  33. (2000). Cytoplasmic beta-catenin accumulation as a predictor of hematogenous metastasis in human colorectal cancer.
  34. (2005). Deregulated NOTCH signaling in acute T-cell lymphoblastic leukemiaAymphoma: New insights, questions, and opportunities.
  35. (1994). Diagnosis of pancreatic cancer and prediction of unresectability using the tumor-associated antigen CA19-9. Pancreas,
  36. (2000). Distinct pattern of expression of differentiation and growth-related genes in squamous cell carcinomas of the head and neck revealed by the use of laser capture microdissection and cDNA arrays.
  37. (2005). Dominant-negative Notch3 receptor inhibits mitogen-activated protein kinase pathway and the growth of human lung cancers. Cancer Res,
  38. (2003). Dysregulation of beta-catenin expression correlates with tumor differentiation in pancreatic duct adenocarcinoma.
  39. (2001). Epidermal growth factor induces cyclin D1 in human pancreatic carcinoma: evidence for a cyclin DII-dependent cell cycle progression.
  40. (1999). Evaluation of cyclin DI mRNA expression in gastric and colorectal cancers. Res Commun Mol Pathol Pharmacol,
  41. (1998). Expression and tissue localization of matrix metalloproteinase 7 (matrilysin) in human gastric carcinomas. Implications for vessel invasion and metastasis.
  42. (1999). Expression of cell-cycle regulatory proteins cyclin D1, cyclin E, and their inhibitor p2l WAFl/CIPI in gastric cancer.
  43. (2005). Expression of Notch-1 and its ligands, Delta-like-I and Jagged-1, is critical for glioma cell surVival and proliferation. Cancer Res,
  44. (2000). Expression of nuclear beta-catenin and c-myc is correlated with tumor size but not with proliferative activity of colorectal adenomas.
  45. (2003). Expression status of Pinl and cyclins in oral squamous cell carcinoma: Pinl correlates with Cyclin D1 mRNA expression and clinical significance of cyclins.
  46. (1995). Food habits and pancreatic cancer: a case-control study of the Francophone community in Montreal, Canada. Cancer EpIdemiol Biomarkers Prev,
  47. (2001). Functional gamma-secretase inhibitors reduce beta-amyloid peptide levels in brain.
  48. (1998). Genetics of pancreatic cancer. From genes to families.
  49. (2002). Glycogen synthase kinase-3beta modulates notch signaling and stability.
  50. (1998). Glycogen synthase kinase3beta regulates cyclin DI proteolysis and subcellular localization.
  51. (1998). Identification of c-MYC as a target of the APC pathway.
  52. Identification of differentially expressed genes in pancreatic cancer cells using cDNA microarray. Cancer Res,
  53. (2004). Identification of the role of presenilins beyond Alzheimees disease.
  54. (1999). Immunophenotypic and prognostic analysis of E-cadherin and beta-catenin expression during breast carcinogenesis and turnour progression: a comparative study with CD44.
  55. (1997). Improvements in survival and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer a randomized trial.
  56. (1998). Increased cyclin 131 in human pancreatic cancer is associated with decreased postoperative survival.
  57. (2001). Induction of cyclin Dl transcription and CDK2 activity by Notch(ic): implication for cell cycle disruption in transformation by Notch(ic). Mol Cell Biol,
  58. (1998). Infrequent expression of p2l is related to altered p53 protein in pancreatic carcinoma.
  59. (1998). Inhibition of basal and mitogen-stimulated pancreatic cancer cell growth by cyclin D1 antisense is associated with loss of turnorigenicity and potentiation of cytotoxicity to cisplatinum.
  60. (1996). InteracUon between Wingless and Notch signaling pathways mediated by dishevelled.
  61. Interrelation between expression of matrix metal loproteinase 7 and beta-catenin in esophageal cancer.
  62. (1997). Ki-ras mutations in exocrine pancreatic cancer: association with clinico-pathological characteristics and with tobacco and alcohol consumption. PANK-ras I Project Investigators.
  63. (1997). Ki-ras mutations in patients with chronic Chapter 6. References pancreatitis: High prevalence but undetermined relevance.
  64. (2002). Loss of Pinl function in the mouse causes phenotypes resembling cyclin DI-null phenotypes.
  65. (2002). Matdx metalloproteinase-7 expression in colorectal cancer liver metastases: evidence for involvement of MMP-7 activation in human cancer metastases. Clin Cancer Res,
  66. (2002). Matrix metal loproteinase-7 is expressed by pancreatic cancer precursors Chapter 6. References and regulates acinar-to-ductal metaplasla in exocrine pancreas.
  67. mechanism and prognostic value of activated AKT in pancreas cancer.
  68. (2002). Molecular causes of colon cancer.
  69. (1988). Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes.
  70. (2004). Multiple niches for Notch in cancer: context is everything.
  71. (2003). Mutation of beta-catenin and its protein accumulation in solid and cystic tumor of the pancreas associated with metastasis.
  72. (1990). Mutational activation of the c-K-ras gene in human pancreatic carcinoma. Badfieres Clin Gastroenterol,
  73. (1997). Neoplastic transformation by truncated alleles of human NOTCH I frAN I and NOTCH2. Mol Cell Blol,
  74. (1999). Notch and wingless regulate expression of cuticle patterning genes. Mol Ce/I Biol,
  75. (2003). Notch mediates TGF alpha-induced changes in epithelial differentiation during pancreatic tumorigenesis.
  76. (2002). Notch receptor cleavage depends on but is not directly executed by presenilins.
  77. (2003). Notch signaling controls multiple steps of pancreatic differentiation.
  78. (2001). Notch signaling in mammary gland tumorigenesis.
  79. (2005). Notch signals control the fate of immature progenitor cells in the intestine.
  80. (2001). Notch-1 regulates NF-kappaB activity in hernopoietic progenitor cells.
  81. (2005). Notch/gamma-secretase inhibition turns proliferative cells in intestinal crypts and adenomas into goblet cells.
  82. (2000). Novel association of a diverse range of genes Wth renal cell carcinoma as identified by differential display.
  83. (2001). Nuclear localization of beta-catenin and plakoglobin in primary and metastatic human colonic carcinomas, colonic adenomas, and normal colon.
  84. (1997). Overexpression of cyclin DI in human pancreatic carcinoma is associated vvith poor prognosis. Cancer Res,
  85. (1999). Overexpression of cyclin Dl occurs in both squamous carcinomas and adenocarcinomas of the esophagus and in adenocarcinomas of the stomach. Hum Pathol,
  86. (2002). Overexpression of Wnt target genes in adenomas of familial adenomatous polyposis patients. Anticancer Res,
  87. (2003). p53 alteration is not an independent prognostic indicator, but affects the efficacy of adjuvant chemotherapy in human pancreatic cancer.
  88. (1995). Pancreaticocluodenectomy for cancer of the head of the pancreas. 201 patients. Ann Surg,
  89. (2003). Phosphorylated Akt/PKB controls cell growth and apoptosis in intraductal papillary-mucinous tumor and invasive ductal adenocarcinoma of the pancreas.
  90. (2001). PinI is overexpressed in breast cancer and cooperates With Ras signaling in increasing the transcriptional activity of c-Jun towards cyclin D1.
  91. (2002). Pinning down proline-directed phosphorylation signaling.
  92. (1999). Prognostic factors associated with resectable adenocarcinoma of the head of the pancreas.
  93. (2004). Prognostic relevance of activated At kinase in node-negative breast cancer: a clinicopathological study of 99 cases. Modem Pathology,
  94. (2004). Prognostic significance of activated Akt expression in pancreatic ductal adenocarcinoma. Clin Cancer Res,
  95. (2002). Prognostic significance of c-myc and AlB1 amplification in hepatocellular carcinoma. A broad survey using high-throughput tissue microarray.
  96. (2002). Prognostic significance of matrix metalloproteinase-7 (MMP-7) expression at the invasive front in gastric carcinoma.
  97. (1998). Prognostic significance of molecular alterations in human pancreatic carcinoma--an immunohistological study. Langenbecks Arch Surg,
  98. (2000). Ras pathway signals are required for notch-mediated oncogenesis.
  99. (2001). Reduced membranous and ectopic cytoplasmic expression of beta -catenin correlate with cyclin D1 overexpression and poor prognosis in pancreatic cancer.
  100. (2001). regulates turnover and subcellular localization of beta- catenin by inhibiting its interaction with APC.
  101. (2003). Relationship between expression of E-cadherin-catenin complex and clinicopathologic characteristics of pancreatic cancer.
  102. (2002). Restoration of E-cadherin/beta-catenin expression in pancreatic cancer cells inhibits growth by induction of apoptosis.
  103. (2001). Results Inhibition of Notch3 using siRNA Chapter 6. References
  104. (1998). Review article: current practice and future perspectives in detection and diagnosis of pancreatic cancer. Aliment Pharmacol Ther,
  105. (1990). Risk of pancreatic cancer in relation to medical history and the use of tobacco, alcohol and coffee.
  106. (2003). Role of NF-kappaB and Akt/Pl3K in the resistance of pancreatic carcinoma cell lines against gemcitabine-induced cell death.
  107. (2003). Role of PTEN and MMP-7 expression in growth, invasion, metastasis and angiogenesis of gastdc carcinoma.
  108. (2004). Role of the phosphatidylinositol 3-kinase-Akt signal pathway in the proliferation of human pancreatic ductal carcinoma cell lines.
  109. Sequence-specific and phosphorylation-dependent proline isomerization: A potential mitotic regulatory mechanism.
  110. (2002). Specific down-modulation of Notchl signaling in cervical cancer cells is required for sustained HPV-E6/E7 expression and late steps of malignant transformation.
  111. (1997). Suppression of Ki-ras p2l levels leading to growth inhibition of pancreatic cancer cell lines with Ki-ras mutation but not those without Ki-ras mutation. Molecular Carcinogenesis,
  112. (2002). The APC gene in colorectal cancer.
  113. (1998). The EnVislon++ system: a new immunohistochernical method for diagnostics and research.
  114. (1998). The essential mitotic peptidyl-prolyl isomerase Pinl binds and regulates mitosis-specific phosphoproteins.
  115. (1997). The link between integration and expression of human papillornavirus type 16 genomes and cellular changes in the evolution of cervical intraepithelial neoplastic lesions.
  116. (1999). The metalloproteinase matrilysin is a target of betacatenin transactivation in intestinal tumors.
  117. (2005). The Notch signaling pathway is related to neurovascular progression of pancreatic cancer. Ann Surg,
  118. (2004). The PI 3-kinase/Akt signaling pathway is activated due to aberrant Pten expression and Chapter 6. References targets transcription factors NF-kappaB and c-Myc in pancreatic cancer cells.
  119. (2003). The prolyl isomerase PinI in breast development and cancer.
  120. (2002). The prolyl isomerase Pinl is a regulator of p53 in genotoxic response.
  121. Therapy of locally unresectable pancreatic carcinoma: a randomized comparison of high dose (6000 rads) radiation alone, moderate dose radiation (4000 rads + 5-fluorouracil), and high dose radiation + 5-fluorouracil: The Gastrointestinal Tumor Study Group.
  122. (2003). Tissue microarray analysis of signal transducers and activators of transcription 3 (Stat3) and Phospho-Stat3 (Tyr7O5) in node- negative breast cancer shows nuclear localization is associated with a better prognosis.
  123. (2004). Toward Alzheimer therapies based on genetic knowledge.
  124. (2003). Upregulation of the oncogene c-myc in Barrett's adenocarcinoma: induction of c-myc by acidified bile acid in vitro.
  125. (2001). Where Notch and Wnt signaling meet. The presenilin hub.

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