title: Detection of E6-AP as a potential therapeutic target in cervical specimen of HPV-infected women.
abstract: OBJECTIVE
  Silencing of HPV oncogenes or their human host client proteins using topically applied small interfering RNA (siRNA) may be an attractive nonsurgical strategy for CIN treatment. An exploratory clinical investigation was designed to evaluate E6-AP mRNA expression levels in different stages of cervical intraepithelial neoplasia and during the menstrual cycle.
  
  
  METHODS
  In 38 premenopausal women aged 18-45 years referred to colposcopy clinic, analysis of serum hormones, cervical smears for cytology and HPV DNA, cervical biopsy, p16 immunohistochemistry and E6-AP mRNA expression levels in cervical smears and biopsies were performed. The intra-subject variability in E6-AP mRNA expression of vaginal smears was assessed and compared to cervical biopsy specimens.
  
  
  RESULTS
  RNA of sufficient quantity and quality was available for E6-AP expression analysis from 97 % of the collected cervical smears and from 56 % of the collected biopsy samples. The normalized RNA levels from cervical smears were approximately tenfold higher compared to biopsies. There was little influence by the phase of the menstrual cycle or by CIN stage. Real-time PCR showed that the expression level of E6-AP is in a range (<28 C t) that would allow for detection of at least 100-fold modulation by a therapeutic agent (based on an assay LOD of C t = 36).
  
  
  CONCLUSIONS
  Our findings suggest a potential therapeutic approach using E6-AP siRNA as a specific molecular target in cervical intraepithelial neoplasia.
authors.affiliation: Department of Obstetrics and Gynecology, Medical University Hannover, Carl-Neuberg-Str. 1, 30625, Hannover, Germany, hillemanns.peter@mh-hannover.de.
authors.forename: Peter
authors.initials: P
authors.lastname: Hillemanns
authors.forename: Matthias
authors.initials: M
authors.lastname: Jentschke
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Evans
authors.forename: Philipp
authors.initials: P
authors.lastname: Soergel
authors.forename: Ralf
authors.initials: R
authors.lastname: Hass
mesh_terms: D000328:Adult; D001707:Biopsy, Needle; D003430:Cross-Sectional Studies; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008597:Menstrual Cycle; D008875:Middle Aged; D030361:Papillomavirus Infections / Q000150:complications*; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D044767:Ubiquitin-Protein Ligases / Q000235:genetics* / Q000378:metabolism; D002583:Uterine Cervical Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D014626:Vaginal Smears; D055815:Young Adult; D002578:Uterine Cervical Dysplasia / Q000378:metabolism* / Q000473:pathology / Q000821:virology
chemical_list: D004279:DNA, Viral; D012333:RNA, Messenger; C471639:UBE3A protein, human; D044767:Ubiquitin-Protein Ligases