The Nuclear Factor-κB RelA Transcription Factor Is Constitutively Activated in Human Pancreatic Adenocarcinoma Cells

Pancreatic adenocarcinoma is a leading cause of adult cancer mortality in the United States. Recent studies have revealed that point mutation of the K-ras oncogene is a common event in pancreatic cancer, and oncogenesis mediated by Ras may also involve activation of Rel/nuclear factor (NF)-kappa B transcription factors. Furthermore, the c-rel member of Rel/NF-kappa B transcription factor family was first identified as a cellular homologue of the v-rel oncogene, suggesting that other members of the Rel/NF-kappa B family are potentially oncogenes. We therefore investigated the possibility that Rel/NF-kappa B transcription factors are activated in pancreatic cancer. Immunohistochemical analysis, Western blot and Northern blot analysis, electrophoretic mobility shift assays, and chloramphenicol acetyltransferase assays were performed to determine RelA activity in human pancreatic adenocarcinomas and normal tissues and nontumorigenic or tumorigenic cell lines. RelA, the p65 subunit of NF-kappa B, was constitutively activated in approximately 67% (16 of 24) of pancreatic adenocarcinomas but not in normal pancreatic tissues. Constitutive RelA activity was also detected in 9 of 11 human pancreatic tumor cell lines but not in nontumorigenic Syrian golden hamster cell lines. I kappa B alpha, a previously identified NF-kappa B-inducible gene, was overexpressed in human pancreatic tumor tissues and cell lines, and RelA activation could be inhibited by curcumin and dominant-negative mutants of I kappa B alpha, raf, and MEKK1. This is the first report demonstrating constitutive activation of RelA in nonlymphoid human cancer. These data are consistent with the possibility that RelA is constitutively activated by the upstream signaling pathway involving Ras and mitogen-activated protein kinases in pancreatic tumor cells. Constitutive RelA activity may play a key role in pancreatic tumorigenesis through activation of its downstream target genes.

[1]  A. E. Rogers,et al.  Aberrant nuclear factor-kappaB/Rel expression and the pathogenesis of breast cancer. , 1997, The Journal of clinical investigation.

[2]  C. Y. Wang,et al.  Requirement of NF-kappaB activation to suppress p53-independent apoptosis induced by oncogenic Ras. , 1997, Science.

[3]  Mike Rothe,et al.  IκB Kinase-β: NF-κB Activation and Complex Formation with IκB Kinase-α and NIK , 1997 .

[4]  Matthias Mann,et al.  IKK-1 and IKK-2: Cytokine-Activated IκB Kinases Essential for NF-κB Activation , 1997 .

[5]  E. Zandi,et al.  The IκB Kinase Complex (IKK) Contains Two Kinase Subunits, IKKα and IKKβ, Necessary for IκB Phosphorylation and NF-κB Activation , 1997, Cell.

[6]  C. Der,et al.  Oncogenic Ha-Ras-induced Signaling Activates NF-κB Transcriptional Activity, Which Is Required for Cellular Transformation* , 1997, The Journal of Biological Chemistry.

[7]  David M. Rothwarf,et al.  A cytokine-responsive IκB kinase that activates the transcription factor NF-κB , 1997, Nature.

[8]  D. Goeddel,et al.  Identification and Characterization of an IκB Kinase , 1997, Cell.

[9]  S. Leach,et al.  Detection of mutated c-Ki-ras in the bile of patients with pancreatic cancer. , 1997, Anticancer research.

[10]  T. Maniatis,et al.  Activation of the IκBα Kinase Complex by MEKK1, a Kinase of the JNK Pathway , 1997, Cell.

[11]  David Baltimore,et al.  An Essential Role for NF-κB in Preventing TNF-α-Induced Cell Death , 1996, Science.

[12]  Marty W. Mayo,et al.  TNF- and Cancer Therapy-Induced Apoptosis: Potentiation by Inhibition of NF-κB , 1996, Science.

[13]  Seamus J. Martin,et al.  Suppression of TNF-α-Induced Apoptosis by NF-κB , 1996, Science.

[14]  D. White,et al.  Rel/NF-kappaB/IkappaB proteins and cancer. , 1996, Oncogene.

[15]  J. Inoue,et al.  MEK Kinase Is Involved in Tumor Necrosis Factor α-Induced NF-κB Activation and Degradation of IκB-α* , 1996, The Journal of Biological Chemistry.

[16]  Scott E. Kern,et al.  DPC4, A Candidate Tumor Suppressor Gene at Human Chromosome 18q21.1 , 1996, Science.

[17]  E M Schwarz,et al.  Rel/NF-kappa B/I kappa B family: intimate tales of association and dissociation. , 1995, Genes & development.

[18]  B. Aggarwal,et al.  Activation of Transcription Factor NF-κB Is Suppressed by Curcumin (Diferuloylmethane) (*) , 1995, The Journal of Biological Chemistry.

[19]  T. Maniatis,et al.  Signal-induced site-specific phosphorylation targets I kappa B alpha to the ubiquitin-proteasome pathway. , 1995, Genes & development.

[20]  R. Bravo,et al.  Overexpression of RelA in transgenic mouse thymocytes: specific increase in levels of the inhibitor protein I kappa B alpha , 1995, Molecular and cellular biology.

[21]  G. Johnson,et al.  Direct Interaction between Ras and the Kinase Domain of Mitogen-activated Protein Kinase Kinase Kinase (MEKK1) (*) , 1995, The Journal of Biological Chemistry.

[22]  S. Gerstberger,et al.  Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation , 1995, Science.

[23]  M. Karin,et al.  Phosphorylation of I kappa B alpha precedes but is not sufficient for its dissociation from NF-kappa B , 1995, Molecular and cellular biology.

[24]  Jeffrey E. Lee,et al.  Preoperative chemoradiation for adenocarcinoma of the pancreas: M.D. Anderson experience , 1995 .

[25]  R. Kumar,et al.  Inhibitory effect of curcumin on epidermal growth factor receptor kinase activity in A431 cells. , 1994, Biochimica et biophysica acta.

[26]  G L Johnson,et al.  Differential activation of ERK and JNK mitogen-activated protein kinases by Raf-1 and MEKK. , 1994, Science.

[27]  N. Sonenberg,et al.  Disruption of I kappa B alpha regulation by antisense RNA expression leads to malignant transformation. , 1994, Oncogene.

[28]  I. Verma,et al.  Enhanced I kappa B alpha degradation is responsible for constitutive NF-kappa B activity in mature murine B-cell lines , 1994, Molecular and cellular biology.

[29]  I. Verma,et al.  Autoregulation of I kappa B alpha activity. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[30]  T. Tan,et al.  Oncogene activation of HIV-LTR-driven expression via the NF-ϰB binding sites , 1993 .

[31]  A. Baldwin,et al.  Kappa B site-dependent induction of gene expression by diverse inducers of nuclear factor kappa B requires Raf-1. , 1993, The Journal of biological chemistry.

[32]  U. Zabel,et al.  Nuclear uptake control of NF‐kappa B by MAD‐3, an I kappa B protein present in the nucleus. , 1993, The EMBO journal.

[33]  M. Nerenberg,et al.  Ablation of transplanted HTLV-I tax-transformed tumors in mice by antisense inhibition of NF-kappa B. , 1992, Science.

[34]  J. McPherson,et al.  Related subunits of NF-kappa B map to two distinct loci associated with translocations in leukemia, NFKB1 and NFKB2. , 1992, Genomics.

[35]  R. Chaganti,et al.  B cell lymphoma-associated chromosomal translocation involves candidate oncogene lyt-10, homologous to NF-κB p50 , 1991, Cell.

[36]  N. Andrews,et al.  A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. , 1991, Nucleic acids research.

[37]  H. Ohno,et al.  The candidate proto-oncogene bcl-3 is related to genes implicated in cell lineage determination and cell cycle control , 1990, Cell.

[38]  D. Baltimore,et al.  I kappa B: a specific inhibitor of the NF-kappa B transcription factor. , 1988, Science.

[39]  D. Shibata,et al.  Most human carcinomas of the exocrine pancreas contain mutant c-K-ras genes , 1988, Cell.

[40]  H. Bose,,et al.  Expression of the v-rel oncogene in reticuloendotheliosis virus-transformed fibroblasts. , 1988, Virology.

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

[42]  H. Temin,et al.  Activation of oncogenicity of the c-rel proto-oncogene , 1986, Molecular and cellular biology.

[43]  P. Wingo,et al.  Cancer statistics, 1997 , 1997, CA: a cancer journal for clinicians.

[44]  Jeffrey E. Lee,et al.  Preoperative chemoradiation, pancreaticoduodenectomy, and intraoperative radiation therapy for adenocarcinoma of the pancreatic head* , 1996 .

[45]  R. Hruban,et al.  Molecular Biology and the Diagnosis and Treatment of Adenocarcinoma of the Pancreas , 1996 .

[46]  A. Baldwin,et al.  THE NF-κB AND IκB PROTEINS: New Discoveries and Insights , 1996 .

[47]  M. Grilli,et al.  NF-kappa B and Rel: participants in a multiform transcriptional regulatory system. , 1993, International review of cytology.

[48]  Hallin Pa,et al.  MODELS OF PANCREATIC CANCER , 1993 .

[49]  H. Bose,,et al.  Transformation of avian lymphoid cells by reticuloendotheliosis virus. , 1988, Mutation research.

[50]  G. Williams,et al.  Application of a cellular test battery in the decision point approach to carcinogen identification. , 1988, Mutation research.

[51]  D. Longnecker,et al.  Experimental carcinogenesis in the pancreas. , 1984, International review of experimental pathology.