PMID- 23724131
OWN - NLM
STAT- MEDLINE
DCOM- 20140110
LR  - 20231106
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 5
DP  - 2013
TI  - Overexpression of p16(INK4a) in urothelial carcinoma in situ is a marker for 
      MAPK-mediated epithelial-mesenchymal transition but is not related to human 
      papillomavirus infection.
PG  - e65189
LID - 10.1371/journal.pone.0065189 [doi]
LID - e65189
AB  - BACKGROUND: The role of human papillomavirus (HPV) in bladder carcinogenesis 
      remains controversial. Overexpression of p16(INK4a), a surrogate marker for 
      infection with oncogenic HPV in other tumours, has been described for urothelial 
      carcinoma in situ (UCIS). Our goal was therefore to evaluate whether 
      overexpression of p16(INK4a) is associated with HPV infection and to identify 
      mechanisms of p16(INK4a) upregulation in UCIS. MATERIALS AND METHODS: In 60 
      tissue specimens from a total of 45 patients (UCIS and controls), we performed 
      p16(INK4a) immunohistochemistry followed by detection and subclassification of 
      HPV DNA. In a subset of samples, we tested for gene amplification of p16(INK4a) 
      applying fluorescence in situ hybridization (FISH). RAS/MAPK signalling and 
      epithelial-mesenchymal transition (EMT) was assessed using immunohistochemistry. 
      Finally, we transfected urothelial carcinoma cells with KRAS and examined the 
      expression of p16(INK4a) as well as markers of EMT. RESULTS: We found 
      overexpression of p16(INK4a) in 92.6% of UCIS and in all cervical intraepithelial 
      neoplasia (CIN) controls. In contrast, we detected high-risk HPV DNA in 80% of 
      CIN, but none in UCIS. There was no gene amplification of p16(INK4a). High levels 
      of phosphorylated kinases and urokinase plasminogen activator (uPA) and loss of 
      membraneous E-cadherin were detected in UCIS. KRAS transfection of urothelial 
      carcinoma cells led to upregulation of p16(INK4a) and uPA accompanied by loss of 
      E-cadherin that could be inhibited by application of the kinase-inhibitor 
      Sorafenib. CONCLUSIONS: Our results show that overexpression of p16(INK4a) in 
      UCIS is neither associated with HPV infection nor p16(INK4a) gene amplification 
      but is a consequence of enhanced RAS/MAPK signalling that promotes EMT, possibly 
      due to Sorafenib-sensitive paracrine secretion of the EMT activator uPA. These 
      findings might open a novel therapeutic option for localized but aggressive 
      urothelial cancer.
FAU - Steinestel, Julie
AU  - Steinestel J
AD  - Department of Urology, University of Ulm, Ulm, Germany.
FAU - Cronauer, Marcus V
AU  - Cronauer MV
FAU - Muller, Johannes
AU  - Muller J
FAU - Al Ghazal, Andreas
AU  - Al Ghazal A
FAU - Skowronek, Peter
AU  - Skowronek P
FAU - Arndt, Annette
AU  - Arndt A
FAU - Kraft, Klaus
AU  - Kraft K
FAU - Schrader, Mark
AU  - Schrader M
FAU - Schrader, Andres J
AU  - Schrader AJ
FAU - Steinestel, Konrad
AU  - Steinestel K
LA  - eng
PT  - Journal Article
DEP - 20130528
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Cyclin-Dependent Kinase Inhibitor p16)
RN  - 0 (Phenylurea Compounds)
RN  - 25X51I8RD4 (Niacinamide)
RN  - 9ZOQ3TZI87 (Sorafenib)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
SB  - IM
MH  - Aged
MH  - Carcinoma in Situ/enzymology/*pathology/virology
MH  - Cell Line, Tumor
MH  - Cyclin-Dependent Kinase Inhibitor p16/*metabolism
MH  - *Epithelial-Mesenchymal Transition/drug effects
MH  - Female
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Hybridization, Fluorescence
MH  - MAP Kinase Signaling System/drug effects
MH  - Male
MH  - Mitogen-Activated Protein Kinases/*metabolism
MH  - Niacinamide/analogs & derivatives/pharmacology
MH  - Papillomaviridae/drug effects/isolation & purification
MH  - Papillomavirus Infections/enzymology/*pathology/virology
MH  - Phenylurea Compounds/pharmacology
MH  - Sorafenib
MH  - Transfection
MH  - Urokinase-Type Plasminogen Activator/metabolism
MH  - Urothelium/*pathology/virology
MH  - Uterine Cervical Neoplasms/enzymology/pathology/virology
MH  - Uterine Cervical Dysplasia/enzymology/pathology/virology
PMC - PMC3665800
COIS- Competing Interests: The authors read the journal's policy and have the following 
      conflicts: Prof. AJ Schrader receives compensation as a consultant for Bayer 
      Healthcare AG, which manufactures Sorafenib (Nexavar(R)) for clinical application. 
      All other authors have declared that no competing interests exist. This does not 
      alter the authors' adherence to all the PLOS ONE policies on sharing data and 
      materials.
EDAT- 2013/06/01 06:00
MHDA- 2014/01/11 06:00
PMCR- 2013/05/28
CRDT- 2013/06/01 06:00
PHST- 2013/01/21 00:00 [received]
PHST- 2013/04/22 00:00 [accepted]
PHST- 2013/06/01 06:00 [entrez]
PHST- 2013/06/01 06:00 [pubmed]
PHST- 2014/01/11 06:00 [medline]
PHST- 2013/05/28 00:00 [pmc-release]
AID - PONE-D-13-03971 [pii]
AID - 10.1371/journal.pone.0065189 [doi]
PST - epublish
SO  - PLoS One. 2013 May 28;8(5):e65189. doi: 10.1371/journal.pone.0065189. Print 2013.