title: p63 drives invasion in keratinocytes expressing HPV16 E6/E7 genes through regulation of Src-FAK signalling.
abstract: Using microarray information from oro-pharyngeal data sets and results from primary human foreskin keratinocytes (HFK) expressing Human Papilloma Virus (HPV)-16 E6/E7 proteins, we show that p63 expression regulates signalling molecules which initiate cell migration such as Src and focal adhesion kinase (FAK) and induce invasion in 3D-organotypic rafts; a phenotype that can be reversed by depletion of p63. Knockdown of Src or FAK in the invasive cells restored focal adhesion protein paxillin at cell periphery and impaired the cell migration. In addition, specific inhibition of FAK (PF573228) or Src (dasatinib) activities mitigated invasion and attenuated the expression/activity of matrix metalloproteinase 14 (MMP14), a pivotal MMP in the MMP activation cascade. Expression of constitutively active Src in non-invasive HFK expressing E6/E7 proteins upregulated the activity of c-Jun and MMP14, and induced invasion in rafts. Depletion of Src, FAK or AKT in the invasive cells normalised the expression/activity of c-Jun and MMP14, thus implicating the Src-FAK/AKT/AP-1 signalling in MMP14-mediated extra-cellular matrix remodelling. Up-regulation of Src, AP-1, MMP14 and p63 expression was confirmed in oro-pharyngeal cancer. Since p63 transcriptionally regulated expression of many of the genes in this signalling pathway, it suggests that it has a central role in cancer progression.
authors.affiliation: Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast BT9 7BL, UK.
authors.forename: Kirtiman
authors.initials: K
authors.lastname: Srivastava
authors.affiliation: Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast BT9 7BL, UK.
authors.forename: Adam
authors.initials: A
authors.lastname: Pickard
authors.affiliation: Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast BT9 7BL, UK.
authors.forename: Simon
authors.initials: S
authors.lastname: McDade
authors.affiliation: Centre for Cancer Research and Cell Biology, Queen's University Belfast, Belfast BT9 7BL, UK.|Department of Pathology, School of Medicine, University of New Mexico, Albuquerque, NM 87131-0001, USA.
authors.forename: Dennis J
authors.initials: DJ
authors.lastname: McCance
mesh_terms: D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism / Q000821:virology; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D000069439:Dasatinib / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D051417:Focal Adhesion Kinase 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D052816:Foreskin / Q000166:cytology; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D008297:Male; D053511:Matrix Metalloproteinase 14 / Q000235:genetics / Q000378:metabolism; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism*; D009959:Oropharyngeal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000821:virology; D050725:Papillomavirus E7 Proteins / Q000235:genetics / Q000378:metabolism*; D030361:Papillomavirus Infections / Q000235:genetics / Q000378:metabolism / Q000821:virology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism; D015363:Quinolones / Q000494:pharmacology; D034622:RNA Interference; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D013450:Sulfones / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: C521108:6-(4-(3-(methylsulfonyl)benzylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydroquinolin-2(1H)-one; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D047428:Protein Kinase Inhibitors; D016755:Proto-Oncogene Proteins c-jun; D015363:Quinolones; D012097:Repressor Proteins; D013450:Sulfones; C115051:TP63 protein, human; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D051417:Focal Adhesion Kinase 1; C496050:PTK2 protein, human; D019061:src-Family Kinases; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14; D000069439:Dasatinib
keywords: HPV16; MMP14; Src; invasion; p63