Adenocarcinoma the Malignant Potential in Human Pancreatic Ductal Converting Enzyme Increases α 17 / Tumor Necrosis Factor-Aberrant Expression of a Disintegrin and Metalloproteinase

A disintegrin and metalloproteinase (ADAM) molecules are known for their unique potential to combine adhesion, proteolysis, and signaling. To understand the role of ADAM17/tumor necrosis factor-A (TNF-A) converting enzyme (TACE) in pancreatic ductal adenocarcinoma (PDAC), we investigated its expression, function, and in vitro regulation. ADAM17/TACE mRNA was expressed in 3 of 10 normal pancreatic tissues, 6 of 8 samples from patients with chronic pancreatitis, 10 of 10 PDAC tissues, and 9 of 9 pancreatic cancer cell lines, but it was absent in primary duct epithelial cells. Immunohistochemical staining revealed positive cancer cells in 8 of 10 PDACs but no staining of ducts in normal pancreas. ADAM17/TACE was found in 0 of 16 pancreatic intraepithelial neoplasia (PanIN)-1A lesions, 1 of 30 PanIN-1B lesions, 2 of 13 PanIN-2 lesions but, in 13 of 15 PanIN-3 lesions, associated with PDAC. Western blot, flow cytometry, and confocal microscopy analyses showed the aberrant expression of ADAM17/TACE protein in pancreatic cancer cell lines. The proteolytic activity of ADAM17/TACE, assessed by the release of TNF-A, was inhibited by TNF-A protease inhibitor. ADAM17/TACE gene silencing using small interfering RNA technique in vitro reduced invasion behavior dramatically, whereas proliferation was unaffected. Furthermore, ADAM17/TACE mRNA expression was down-regulated in pancreatic cancer cells arrested in G2-M phase as well as in a time-dependent manner after TNF-A and interleukin-6 incubation. In conclusion, our findings provide evidence of aberrant expression of the proteolytically active ADAM17/TACE in advanced precursor lesions (PanIN-3) and PDAC while identifying its critical involvement in the invasion process. (Cancer Res 2006; 66(18): 9045-53)

[1]  G. Weskamp,et al.  Distinct roles for ADAM10 and ADAM17 in ectodomain shedding of six EGFR ligands , 2004, The Journal of cell biology.

[2]  H. Samaratunga,et al.  Expression of the Disintegrin Metalloprotease, ADAM-10, in Prostate Cancer and Its Regulation by Dihydrotestosterone, Insulin-Like Growth Factor I, and Epidermal Growth Factor in the Prostate Cancer Cell Model LNCaP , 2004, Clinical Cancer Research.

[3]  S. Batra,et al.  Expression of ADAMs (a disintegrin and metalloproteases) and TIMP-3 (tissue inhibitor of metalloproteinase-3) in human prostatic adenocarcinomas. , 2003, International journal of oncology.

[4]  J. White.  ADAMs: modulators of cell-cell and cell-matrix interactions. , 2003, Current opinion in cell biology.

[5]  M. W. Büchler,et al.  Microarray-based identification of differentially expressed growth- and metastasis-associated genes in pancreatic cancer , 2003, Cellular and Molecular Life Sciences CMLS.

[6]  Stefan Hart,et al.  TACE cleavage of proamphiregulin regulates GPCR‐induced proliferation and motility of cancer cells , 2003, The EMBO journal.

[7]  H. Friess,et al.  Mucin (MUC) gene expression in human pancreatic adenocarcinoma and chronic pancreatitis: a potential role of MUC4 as a tumor marker of diagnostic significance. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[8]  G. Zamboni,et al.  The Immunohistochemical Mucin Expression Pattern Distinguishes Different Types of Intraductal Papillary Mucinous Neoplasms of the Pancreas and Determines Their Relationship to Mucinous Noncystic Carcinoma and Ductal Adenocarcinoma , 2001, The American journal of surgical pathology.

[9]  R H Hruban,et al.  Pancreatic Intraepithelial Neoplasia: A New Nomenclature and Classification System for Pancreatic Duct Lesions , 2001, The American journal of surgical pathology.

[10]  C. Blobel,et al.  Remarkable roles of proteolysis on and beyond the cell surface. , 2000, Current opinion in cell biology.

[11]  D. McCulloch,et al.  The expression of the ADAMs proteases in prostate cancer cell lines and their regulation by dihydrotestosterone , 2000, Molecular and Cellular Endocrinology.

[12]  Y. DeClerck.  Interactions between tumour cells and stromal cells and proteolytic modification of the extracellular matrix by metalloproteinases in cancer. , 2000, European journal of cancer.

[13]  S. Rose-John,et al.  Shedding of interleukin-6 receptor and tumor necrosis factor alpha. Contribution of the stalk sequence to the cleavage pattern of transmembrane proteins. , 2000, European journal of biochemistry.

[14]  J. Buxbaum,et al.  Tumor Necrosis Factor-α-converting Enzyme Is Required for Cleavage of erbB4/HER4* , 2000, The Journal of Biological Chemistry.

[15]  C. Blobel,et al.  Intracellular maturation and localization of the tumour necrosis factor alpha convertase (TACE). , 2000, The Biochemical journal.

[16]  R H Hruban,et al.  Hypermethylation of multiple genes in pancreatic adenocarcinoma. , 2000, Cancer research.

[17]  C. Blobel,et al.  Evidence for an interaction of the metalloprotease-disintegrin tumour necrosis factor alpha convertase (TACE) with mitotic arrest deficient 2 (MAD2), and of the metalloprotease-disintegrin MDC9 with a novel MAD2-related protein, MAD2beta. , 1999, The Biochemical journal.

[18]  G. Wu,et al.  Cutting edge: a dominant negative form of TNF-alpha converting enzyme inhibits proTNF and TNFRII secretion. , 1999, Journal of immunology.

[19]  Fengzhi Li,et al.  Control of apoptosis and mitotic spindle checkpoint by survivin , 1998, Nature.

[20]  David C. Lee,et al.  An essential role for ectodomain shedding in mammalian development. , 1998, Science.

[21]  R. Black,et al.  ADAMs: focus on the protease domain. , 1998, Current opinion in cell biology.

[22]  M. Löhr,et al.  Increasing the Transfection Efficacy and Subsequent Long‐Term Culture of Resting Human Pancreatic Duct Epithelial Cells , 1998, Pancreas.

[23]  M. T. Brewer,et al.  Identification and Characterization of a Pro-tumor Necrosis Factor-α-processing Enzyme from the ADAM Family of Zinc Metalloproteases* , 1997, The Journal of Biological Chemistry.

[24]  P. Croucher,et al.  Expression of members of the novel membrane linked metalloproteinase family ADAM in cells derived from a range of haematological malignancies. , 1997, Biochemical and biophysical research communications.

[25]  M. Löhr,et al.  Expression and Function of Receptors for Extracellular Matrix Proteins in Human Ductal Adenocarcinomas of the Pancreas , 1996, Pancreas.

[26]  W. Telford,et al.  Evaluation of glucocorticoid‐induced DNA fragmentation in mouse thymocytes by flow cytometry , 1991, Cell proliferation.

[27]  M. Korc.  Potential role of the epidermal growth factor receptor in human pancreatic cancer , 1990, International journal of pancreatology : official journal of the International Association of Pancreatology.

[28]  R. Metzgar,et al.  Characterization of Clones of a Human Pancreatic Adenocarcinoma Cell Line Representing Different Stages of Differentiation , 1989, Pancreas.

[29]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[30]  G. Koch,et al.  Synchronization of HeLa cell cultures by inhibition of DNA polymerase alpha with aphidicolin. , 1980, Nucleic acids research.

[31]  R. Maciewicz,et al.  Integrin alpha5beta1 and ADAM-17 interact in vitro and co-localize in migrating HeLa cells. , 2004, The Journal of biological chemistry.

[32]  H. Friess,et al.  Pancreatic cancer: the potential clinical relevance of alterations in growth factors and their receptors , 2004, Journal of Molecular Medicine.

[33]  D. Seals,et al.  The ADAMs family of metalloproteases: multidomain proteins with multiple functions. , 2003, Genes & development.

[34]  J. Baselga,et al.  TACE is required for the activation of the EGFR by TGF-alpha in tumors. , 2003, The EMBO journal.

[35]  C. Blobel,et al.  Tumor necrosis factor-alpha converting enzyme/ADAM 17 mediates MUC1 shedding. , 2003, The Journal of biological chemistry.

[36]  J. Ringel,et al.  Transforming Growth Factor-β1 Induces Desmoplasia in an Experimental Model of Human Pancreatic Carcinoma , 2001 .

[37]  R. Black,et al.  Functional analysis of the domain structure of tumor necrosis factor-alpha converting enzyme. , 2000, The Journal of biological chemistry.

[38]  F. Loechel,et al.  Cysteine-rich domain of human ADAM 12 (meltrin alpha) supports tumor cell adhesion. , 1999, The American journal of pathology.

[39]  L. Lum,et al.  Evidence for a role of a tumor necrosis factor-alpha (TNF-alpha)-converting enzyme-like protease in shedding of TRANCE, a TNF family member involved in osteoclastogenesis and dendritic cell survival. , 1999, The Journal of biological chemistry.

[40]  P. Slocombe,et al.  TNF-alpha converting enzyme (TACE) is inhibited by TIMP-3. , 1998, FEBS letters.

[41]  B. Castner,et al.  A metalloproteinase disintegrin that releases tumour-necrosis factor-alpha from cells. , 1997, Nature.

[42]  M. Lambert,et al.  Cloning of a disintegrin metalloproteinase that processes precursor tumour-necrosis factor-α , 1997, Nature.

[43]  U. Rodeck,et al.  Tumor necrosis factor a induces the expression of transforming growth factor a and the epidermal growth factor receptor in human pancreatic cancer cells , 2022 .