381 ABERRANT EXTRACELLULAR SIGNAL-REGULATED PROTEIN KINASE 5 (ERK5) SIGNALLING PROMOTES CELLULAR MOTILITY AND INVASION IN PROSTATE CARCINOGENESIS

Abstract

You have accessJournal of UrologyProstate Cancer: Basic Research III1 Apr 2010381 ABERRANT EXTRACELLULAR SIGNAL-REGULATED PROTEIN KINASE 5 (ERK5) SIGNALLING PROMOTES CELLULAR MOTILITY AND INVASION IN PROSTATE CARCINOGENESIS Alison Ramsay, Stuart McCracken, Rosie Morland, Janis Fleming, Laura Machesky, Xinzi Yu, Dylan Edwards, Robert Nuttall, Morag Seywright, Evan Keller, and Hing Leung Alison RamsayAlison Ramsay Glasgow, United Kingdom More articles by this author , Stuart McCrackenStuart McCracken Newcastle Upon Tyne, United Kingdom More articles by this author , Rosie MorlandRosie Morland Glasgow, United Kingdom More articles by this author , Janis FlemingJanis Fleming Glasgow, United Kingdom More articles by this author , Laura MacheskyLaura Machesky Glasgow, United Kingdom More articles by this author , Xinzi YuXinzi Yu Glasgow, United Kingdom More articles by this author , Dylan EdwardsDylan Edwards Norwich, United Kingdom More articles by this author , Robert NuttallRobert Nuttall Norwich, United Kingdom More articles by this author , Morag SeywrightMorag Seywright Glasgow, United Kingdom More articles by this author , Evan KellerEvan Keller Ann Arbor, MI More articles by this author , and Hing LeungHing Leung Glasgow, United Kingdom More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2010.02.449AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The MEK5/ERK5 pathway is implicated in a number of tumour types including prostate cancer. Its molecular mechanism for carcinogenesis remains to be fully characterised. We have further investigated the functional role of ERK5 in prostate cancer and tested its relationship ERK1/2. METHODS Functional assays were performed in prostate cancer cells (PC3 and PC3M cells) following transfection with ERK1, ERK2 and ERK5 siRNA or treatment with the mitogen-activated protein kinase kinase (MEK) inhibitor (PD184352): proliferation by cell count, motility by culture chamber, migration/invasion by time-lapse microscropy. Matrix metalloproteinase (MMP) expression was studied by promoter luciferase reporter assay and real time PCR. Invadopodia assays were performed as described previously (J Cell Sci 2008, 121: 369), and data were analysed using Image J software. Immunohistochemistry for ERK5 expression was also performed on metastatic prostate tissue (Oncogene 2008, 27: 2978). RESULTS Using siRNA and PD184352 to target ERK5 expression and function respectively, both motility and invasion of PC3 cells were significantly impaired (P<0.005 for all parameters). Q-PCR and promoter activity analysis revealed activation of MMP-2, -9, -12 and TIMP2 expression, and suppression of MMP-16 by ERK5. Following ERK5 transfection, PC3 cells formed significantly more invadopodia (P=0.0048). Immunohistochemistry further confirmed upregulated ERK5 expression in both primary and metastatic prostate tumours. Among the informative metastatic lesions (n=11), strong ERK5 immunoreactivity, cytoplasmic (55%) and nuclear (73%), was observed. Potential ‘cross-talk' between ERK5 and ERK1/2 was explored. ERK1 knockdown resulted in prolonged and enhanced activation of both ERK5 and ERK2. This was associated with a small but reproducible mitogenic effect following ERK1 specific knockdown, supporting published data that ERK1 acts as a negative regulator and ERK2 as a positive regulator of cell proliferation. CONCLUSIONS Our results validate the importance of the ERK5 signalling pathway as a potential target for therapy and highlight a novel functional and biochemical relationship between ERK1 and ERK5 signalling. © 2010 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 183Issue 4SApril 2010Page: e151 Advertisement Copyright & Permissions© 2010 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alison Ramsay Glasgow, United Kingdom More articles by this author Stuart McCracken Newcastle Upon Tyne, United Kingdom More articles by this author Rosie Morland Glasgow, United Kingdom More articles by this author Janis Fleming Glasgow, United Kingdom More articles by this author Laura Machesky Glasgow, United Kingdom More articles by this author Xinzi Yu Glasgow, United Kingdom More articles by this author Dylan Edwards Norwich, United Kingdom More articles by this author Robert Nuttall Norwich, United Kingdom More articles by this author Morag Seywright Glasgow, United Kingdom More articles by this author Evan Keller Ann Arbor, MI More articles by this author Hing Leung Glasgow, United Kingdom More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...