Abstract
The recognition of biologically distinct tumor subsets is fundamental to understanding tumorigenesis. This study investigated the mutational status of the serine/threonine kinase BRAF and the cyclin E regulator FBXW7 (CDC4, FBW7, AGO, SEL10) related to two distinct pancreatic carcinoma subsets: the medullary KRAS2-wild-type and the cyclin E overexpressing tumors, respectively. Among KRAS2-wild-type carcinomas, 33% (3 of 9) contained BRAF V599E mutations; one of which was identified in the pancreatic cancer cell line COLO357. Among 74 KRAS2-mutant carcinomas, no BRAF mutations were identified. Among the KRAS2/BRAF wild-type carcinomas, no mutations within pathway members MEK1, MEK2, ERK1, ERK2, RAP1B, or BAD were found. Using pancreatic cancer microarrays and immunohistochemistry, we determined that 6% (4 of 46 and 5 of 100 in two independent panels) of pancreatic adenocarcinomas overexpress cyclin E. We identified two potential mechanisms for this overexpression including the amplification/gain of CCNE1 gene copies in the Panc-1 and Su86.86 cell lines and a novel somatic homozygous mutation (H460R, in one of 11 pancreatic cancer xenografts having allelic loss) in FBXW7, which was accompanied by cyclin E overexpression by immunohistochemistry. Both BRAF and FBXW7 mutations functionally activate kinase effectors important in pancreatic cancer and extend the potential options for therapeutic targeting of kinases in the treatment of phenotypically distinct pancreatic adenocarcinoma subsets.
MeSH Terms
Adenocarcinoma/genetics,metabolism,therapy
Amino Acid Sequence/genetics
Animals
Base Sequence/genetics
Cell Cycle Proteins/genetics
Cyclin E/genetics,metabolism
F-Box Proteins
F-Box-WD Repeat-Containing Protein 7
Gene Amplification
Gene Dosage
Humans
Immunohistochemistry
Mice
Molecular Sequence Data
Mutation
Oncogene Proteins/genetics
Pancreatic Neoplasms/genetics,metabolism,therapy
Proto-Oncogene Proteins B-raf
Tumor Cells, Cultured
Ubiquitin-Protein Ligases
Chemicals
Cell Cycle Proteins
Cyclin E
F-Box Proteins
F-Box-WD Repeat-Containing Protein 7
FBXW7 protein, human
Fbxw7 protein, mouse
Oncogene Proteins
Ubiquitin-Protein Ligases
BRAF protein, human
Braf protein, mouse
Proto-Oncogene Proteins B-raf
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Calhoun Eric S
Department of Oncology, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Baltimore, Maryland 21231, USA.
Jones Jessa B
Ashfaq Raheela
Adsay Volkan
Baker Suzanne J
Valentine Virginia
Hempen Paula M
Hilgers Werner
Yeo Charles J
Hruban Ralph H
Kern Scott E
References (27)
27 references, click to expand
-
Genetic profile of gliosarcomas.
Am J Pathol. 2000 Feb;156(2):425-32
PMID: 10666371
-
Frequent somatic mutations and homozygous deletions of the p16 (MTS1) gene in pancreatic adenocarcinoma.
Nat Genet. 1994 Sep;8(1):27-32
PMID: 7726912
-
High-throughput tissue microarray analysis of cyclin E gene amplification and overexpression in urinary bladder cancer.
Am J Pathol. 2000 Sep;157(3):787-94
PMID: 10980118
-
Identification and characterization of a 19q12 amplicon in esophageal adenocarcinomas reveals cyclin E as the best candidate gene for this amplicon.
Cancer Res. 2000 Dec 15;60(24):7021-7
PMID: 11156406
-
Reciprocal expressions of cyclin E and cyclin D1 in hepatocellular carcinoma.
Cancer Lett. 2001 Jul 10;168(1):57-63
PMID: 11368878
-
Archipelago regulates Cyclin E levels in Drosophila and is mutated in human cancer cell lines.
Nature. 2001 Sep 20;413(6853):311-6
PMID: 11565033
-
Human F-box protein hCdc4 targets cyclin E for proteolysis and is mutated in a breast cancer cell line.
Nature. 2001 Sep 20;413(6853):316-22
PMID: 11565034
-
Phosphorylation-dependent ubiquitination of cyclin E by the SCFFbw7 ubiquitin ligase.
Science. 2001 Oct 5;294(5540):173-7
PMID: 11533444
-
APC/CTNNB1 (beta-catenin) pathway alterations in human prostate cancers.
Genes Chromosomes Cancer. 2002 May;34(1):9-16
PMID: 11921277
-
Mutations of the BRAF gene in human cancer.
Nature. 2002 Jun 27;417(6892):949-54
PMID: 12068308
-
Cyclins in breast cancer: too much of a good thing.
Breast Cancer Res. 2002;4(4):145-7
PMID: 12100739
-
Reciprocal Rb inactivation and p16INK4 expression in primary lung cancers and cell lines.
Cancer Res. 1995 Feb 1;55(3):505-9
PMID: 7834618
-
Inactivation of the type II TGF-beta receptor in colon cancer cells with microsatellite instability.
Science. 1995 Jun 2;268(5215):1336-8
PMID: 7761852
-
Amplification of AKT2 in human pancreatic cells and inhibition of AKT2 expression and tumorigenicity by antisense RNA.
Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3636-41
PMID: 8622988
-
Molecular cloning and chromosomal localization of the human cyclin C (CCNC) and cyclin E (CCNE) genes: deletion of the CCNC gene in human tumors.
Genomics. 1996 Mar 1;32(2):253-9
PMID: 8833152
-
Overexpression of MDM2, due to enhanced translation, results in inactivation of wild-type p53 in Burkitt's lymphoma cells.
Oncogene. 1998 Mar 26;16(12):1603-10
PMID: 9569028
-
Pancreatic adenocarcinomas with DNA replication errors (RER+) are associated with wild-type K-ras and characteristic histopathology. Poor differentiation, a syncytial growth pattern, and pushing borders suggest RER+.
Am J Pathol. 1998 Jun;152(6):1501-7
PMID: 9626054
-
Establishment and validation of real-time polymerase chain reaction method for CDH1 promoter methylation.
Am J Pathol. 2002 Aug;161(2):629-34
PMID: 12163387
-
hCDC4 gene mutations in endometrial cancer.
Cancer Res. 2002 Aug 15;62(16):4535-9
PMID: 12183400
-
Tumorigenesis: RAF/RAS oncogenes and mismatch-repair status.
Nature. 2002 Aug 29;418(6901):934
PMID: 12198537
-
PDGFRA activating mutations in gastrointestinal stromal tumors.
Science. 2003 Jan 31;299(5607):708-10
PMID: 12522257
-
Evidence of selection for clones having genetic inactivation of the activin A type II receptor (ACVR2) gene in gastrointestinal cancers.
Cancer Res. 2003 Mar 1;63(5):994-9
PMID: 12615714
-
Mutations in BRAF and KRAS characterize the development of low-grade ovarian serous carcinoma.
J Natl Cancer Inst. 2003 Mar 19;95(6):484-6
PMID: 12644542
-
Inverse regulation of human ERBB2 and epidermal growth factor receptors by tumor necrosis factor alpha.
Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8972-6
PMID: 8105469
-
Alterations in the PITSLRE protein kinase gene complex on chromosome 1p36 in childhood neuroblastoma.
Nat Genet. 1994 Jul;7(3):370-5
PMID: 7920654
-
Absence of p16INK4 protein is restricted to the subset of lung cancer lines that retains wildtype RB.
Oncogene. 1994 Nov;9(11):3375-8
PMID: 7936665
-
Genetic, immunohistochemical, and clinical features of medullary carcinoma of the pancreas: A newly described and characterized entity.
Am J Pathol. 2000 May;156(5):1641-51
PMID: 10793075