Inactivation of the mitogen-activated protein kinase pathway as a potential target-based therapy in ovarian serous tumors with KRAS or BRAF mutations.

Activation of mitogen-activated protein kinase (MAPK) occurs in response to various growth stimulating signals and as a result of activating mutations of the upstream regulators, KRAS and BRAF, which can be found in many types of human cancer. To investigate the roles of MAPK activation in tumors harboring KRAS or BRAF mutations, we inactivated MAPK in ovarian tumor cells using CI-1040, a compound that selectively inhibits MAPK kinase, an upstream regulator of MAPK and thus prevents MAPK activation. Profound growth inhibition and apoptosis were observed in CI-1040-treated tumor cells with mutations in either KRAS or BRAF in comparison with the ovarian cancer cells containing wild-type sequences. Long serial analysis of gene expression identified several differentially expressed genes in CI-1040-treated MPSC1 cells harboring an activating mutation in BRAF (V599L). The most striking changes were down-regulation of cyclin D1, COBRA1, and transglutaminase-2 and up-regulation of tumor necrosis factor-related apoptosis-induced ligand, thrombospondin-1, optineurin, and palladin. These patterns of gene expression were validated in other CI-1040-treated tumor cells based on quantitative PCR. Constitutive expression of cyclin D1 partially reversed the growth inhibitory effect of CI-1040 in MPSC1 cells. Our findings indicate that an activated MAPK pathway is critical in tumor growth and survival of ovarian tumors with KRAS or BRAF mutations and suggest that the CI-1040 induced phenotypes depend on the mutational status of KRAS and BRAF in ovarian tumors.

[1]  J. Sebolt-Leopold,et al.  CI-1040 (PD184352), a targeted signal transduction inhibitor of MEK (MAPKK). , 2003, Seminars in oncology.

[2]  D. Barford,et al.  Mechanism of Activation of the RAF-ERK Signaling Pathway by Oncogenic Mutations of B-RAF , 2004, Cell.

[3]  E. Collisson,et al.  Treatment of metastatic melanoma with an orally available inhibitor of the Ras-Raf-MAPK cascade. , 2003, Cancer research.

[4]  F. Montz,et al.  Micropapillary serous ovarian carcinoma: surgical management and clinical outcome. , 2002, Gynecologic oncology.

[5]  A. Sparks,et al.  Using the transcriptome to annotate the genome , 2002, Nature Biotechnology.

[6]  R. Hynes,et al.  Thrombospondin-1 suppresses spontaneous tumor growth and inhibits activation of matrix metalloproteinase-9 and mobilization of vascular endothelial growth factor , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. Olson,et al.  p38 MAP kinase: a convergence point in cancer therapy. , 2004, Trends in molecular medicine.

[8]  L. Cope,et al.  Patterns of p53 Mutations Separate Ovarian Serous Borderline Tumors and Low- and High-grade Carcinomas and Provide Support for a New Model of Ovarian Carcinogenesis: A Mutational Analysis With Immunohistochemical Correlation , 2005, The American journal of surgical pathology.

[9]  F. Gheyas,et al.  Analysis of the ERK1,2 transcriptome in mammary epithelial cells. , 2004, The Biochemical journal.

[10]  David W. Dawson,et al.  CD36 Mediates the In Vitro Inhibitory Effects of Thrombospondin-1 on Endothelial Cells , 1997, The Journal of cell biology.

[11]  Tian-Li Wang,et al.  Identifying tumor origin using a gene expression-based classification map. , 2003, Cancer research.

[12]  A. Nicholson,et al.  Mutations of the BRAF gene in human cancer , 2002, Nature.

[13]  J. Sebolt-Leopold.  MEK inhibitors: a therapeutic approach to targeting the Ras-MAP kinase pathway in tumors. , 2004, Current pharmaceutical design.

[14]  N. Fusenig,et al.  Tumor suppression in human skin carcinoma cells by chromosome 15 transfer or thrombospondin-1 overexpression through halted tumor vascularization. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[15]  J. Tschopp,et al.  TRAIL receptors 1 (DR4) and 2 (DR5) signal FADD-dependent apoptosis and activate NF-kappaB. , 1997, Immunity.

[16]  James M. Roberts,et al.  CDK inhibitors: positive and negative regulators of G1-phase progression. , 1999, Genes & development.

[17]  O. Volpert,et al.  Id1 regulates angiogenesis through transcriptional repression of thrombospondin-1. , 2002, Cancer cell.

[18]  I. Shih,et al.  Diverse tumorigenic pathways in ovarian serous carcinoma. , 2002, The American journal of pathology.

[19]  Chung-Liang Ho,et al.  Mutations of BRAF and KRAS Precede the Development of Ovarian Serous Borderline Tumors , 2004, Cancer Research.

[20]  Carlos L Arteaga,et al.  Tyrosine kinase inhibitors: why does the current process of clinical development not apply to them? , 2004, Cancer cell.

[21]  Brigitte M. Ronnett,et al.  The Histologic Type and Stage Distribution of Ovarian Carcinomas of Surface Epithelial Origin , 2004, International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists.

[22]  Rita Casadio,et al.  Transglutaminases: nature's biological glues. , 2002, The Biochemical journal.

[23]  Alan R. Saltiel,et al.  Blockade of the MAP kinase pathway suppresses growth of colon tumors in vivo , 1999, Nature Medicine.

[24]  Arul M. Chinnaiyan,et al.  The Receptor for the Cytotoxic Ligand TRAIL , 1997, Science.

[25]  I. Shih,et al.  Mutations in BRAF and KRAS characterize the development of low-grade ovarian serous carcinoma. , 2003, Journal of the National Cancer Institute.

[26]  K. Kinzler,et al.  Tumorigenesis: RAF/RAS oncogenes and mismatch-repair status , 2002, Nature.

[27]  Chung-Liang Ho,et al.  Characterization of Active Mitogen-Activated Protein Kinase in Ovarian Serous Carcinomas , 2004, Clinical Cancer Research.

[28]  B. Ponder,et al.  Overexpression of cyclin D1 in epithelial ovarian cancers. , 1997, Gynecologic oncology.

[29]  S. Cook,et al.  Sustained MAP kinase activation is required for the expression of cyclin D1, p21Cip1 and a subset of AP-1 proteins in CCL39 cells , 1999, Oncogene.

[30]  C. Sherr.  Cell cycle control and cancer. , 2000, Harvey lectures.

[31]  H. Schaeffer,et al.  Mitogen-Activated Protein Kinases: Specific Messages from Ubiquitous Messengers , 1999, Molecular and Cellular Biology.

[32]  Ji Huang,et al.  [Serial analysis of gene expression]. , 2002, Yi chuan = Hereditas.

[33]  R. Weinberg,et al.  Ras modulates Myc activity to repress thrombospondin-1 expression and increase tumor angiogenesis. , 2003, Cancer cell.

[34]  J. Sebolt-Leopold,et al.  Biomarker Assays for Phosphorylated MAP Kinase , 2003 .

[35]  J. Pouysségur,et al.  Cyclin D1 Expression Is Regulated Positively by the p42/p44MAPK and Negatively by the p38/HOGMAPK Pathway* , 1996, The Journal of Biological Chemistry.

[36]  Anne E Carpenter,et al.  BRCA1-induced large-scale chromatin unfolding and allele-specific effects of cancer-predisposing mutations , 2001, The Journal of cell biology.

[37]  M. Skobe,et al.  Overexpression of thrombospondin-1 decreases angiogenesis and inhibits the growth of human cutaneous squamous cell carcinomas. , 1999, The American journal of pathology.

[38]  J. Sebolt-Leopold,et al.  Biomarker assays for phosphorylated MAP kinase. Their utility for measurement of MEK inhibition. , 2003, Methods in molecular medicine.

[39]  F. Montz,et al.  Recurrent micropapillary serous ovarian carcinoma , 2002, Cancer.

[40]  C. Peyssonnaux,et al.  The Raf/MEK/ERK pathway: new concepts of activation , 2001, Biology of the cell.

[41]  A. C. Boyd,et al.  Isolation and computer-aided characterization of MmeI, a type II restriction endonuclease from Methylophilus methylotrophus , 1986, Nucleic Acids Res..

[42]  A. Flanagan,et al.  In ovarian neoplasms, BRAF, but not KRAS, mutations are restricted to low‐grade serous tumours , 2004, The Journal of pathology.

[43]  C A Smith,et al.  Identification and characterization of a new member of the TNF family that induces apoptosis. , 1995, Immunity.