title: Immunostaining of p16(INK4a)/Ki-67 and L1 capsid protein on liquid-based cytology specimens obtained from ASC-H and LSIL-H cases.
abstract: BACKGROUND
  Atypical squamous cell cannot exclude high-grade squamous intraepithelial lesion (ASC-H) and low-grade intraepithelial lesion cannot exclude high-grade squamous intraepithelial lesion (LSIL-H) are ambiguous diagnostic entities for the prediction of high-grade cervical lesion. Objective and reproducible tests for predicting high-grade cervical lesions are needed to reduce unnecessary colposcopic referrals or follow-ups.
  
  
  OBJECTIVE
  We aimed to identify an adequate set of adjunctive markers to predict cervical intraepithelial neoplasia grade 2+ (CIN2+) in residual liquid-based cytology specimens (LBCS).
  
  
  METHODS
  We conducted p16 (INK4a)/Ki-67 and L1 capsid protein immunostaining and human papillomavirus (HPV) DNA typing on 56 LBCS diagnosed with ASC-H or LSIL-H, all of which were subjected to histologic confirmation or follow-up cytologic examination.
  
  
  RESULTS
  Positivity for p16 (INK4a)/Ki-67 was associated with a histology of CIN2+ (P=0.047) and CIN3+ (P=0.002). Negativity for L1 capsid protein was associated with CIN2+ confirmed at follow-up (P=0.02).Positivity for high-risk HPV (HR-HPV) was associated with CIN2+ confirmed at follow-up (P=0.036) and a histology of CIN2+ (P=0.037). The sensitivity, specificity, positive predictive value, and negative predictive value for predicting follow-up CIN2+ were 76.2%, 51.4%, 48.5%, and 78.3%, respectively, for p16 (INK4a)/Ki-67 immunostaining; 95.2%, 34.3%, 46.5%, and 92.3%, respectively, for L1 capsid protein; and 66.7%, 67.7%, 54.5%, and 77.8%, respectively, for HR-HPV. The classification and regression tree analysis showed that the combined results of p16 (INK4a)/Ki-67 andL1 capsid protein immunostaining and the HR-HPV test, conducted sequentially, correctly classified 81.8% of samples (27/33)in the prediction of a histology of CIN2 + in ASC-H or LSIL-H. For determination of the histology of cervical intraepithelial neoplasia grade 3+ (CIN3+)in ASC-H or LSIL-H, we found that the combined results of p16 (INK4a)/Ki-67 and L1 capsid protein immunostaining correctly classified 78.8% (26/33) of samples.
  
  
  CONCLUSIONS
  p16(INK4a)/Ki-67 and L1 capsid protein immunostaining and HR-HPV testing of residual LBCS diagnosed with ASC-H or LSIL-H are useful objective biomarkers for predicting CIN2+. Immunostaining for p16(INK4a)/Ki-67 and L1 capsid protein are sufficient to predict CIN3+.
authors.affiliation: 1. Department of Obstetrics and Gynecology, The Catholic University of Korea, Seoul, Korea;
authors.forename: Seung Won
authors.initials: SW
authors.lastname: Byun
authors.forename: Ahwon
authors.initials: A
authors.lastname: Lee
authors.forename: Suyeon
authors.initials: S
authors.lastname: Kim
authors.forename: Yeong Jin
authors.initials: YJ
authors.lastname: Choi
authors.forename: Youn Soo
authors.initials: YS
authors.lastname: Lee
authors.forename: Jong Sup
authors.initials: JS
authors.lastname: Park
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000235:genetics / Q000302:isolation & purification; D036022:Capsid Proteins / Q000235:genetics / Q000302:isolation & purification*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000302:isolation & purification*; D003581:Cytodiagnosis / Q000379:methods*; D005260:Female; D052162:Human papillomavirus 16 / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D019394:Ki-67 Antigen / Q000235:genetics / Q000302:isolation & purification*; D008875:Middle Aged; D009367:Neoplasm Staging; D011247:Pregnancy; D014626:Vaginal Smears; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis* / Q000473:pathology / Q000821:virology
chemical_list: D014408:Biomarkers, Tumor; D036022:Capsid Proteins; D019941:Cyclin-Dependent Kinase Inhibitor p16; D019394:Ki-67 Antigen
keywords: Cervicovaginal cytology; HPV DNA typing.; Human papillomavirus; Immunocytochemistry; L1 capsid protein; p16 INK4a/Ki-67 dual staining