title: mRNA biomarker detection in liquid-based cytology: a new approach in the prevention of cervical cancer.
abstract: Several high-risk human papillomavirus (HPV)-induced cell biomarkers have been proposed as possible candidates to identify patients harboring high-grade squamous intraepithelial lesions (HSILs) of the uterine cervix. We aimed to determine the feasibility of the detection of the mRNA of six biomarkers in cervical smear specimens obtained by liquid-based cytology and to evaluate whether this approach might be useful in the identification of patients with HSIL. One-hundred and twenty three women referred to colposcopy in the Hospital Clinic of Barcelona were included in the study. After a thorough study, including Pap test, high-risk HPV testing (Hybrid Capture 2 test), and colposcopy with directed biopsy and/or endocervical curettage, 48 patients were diagnosed with HSIL, whereas 75 were classified as negative (n=28), or harboring low-grade SIL (n=47). CDKN2A/p16, BIRC5, MMP9, TOP2A, MCM5, and MKI67 mRNA expression was analyzed by reverse transcription quantitative polymerase chain reaction in liquid-based cytology after the Pap test and Hybrid Capture 2 performance. The tissue expression of these biomarkers was analyzed by immunohistochemistry in the biopsy material. One-hundred and thirteen out of 123 (92%) liquid-based cytology yielded adequate material for mRNA analysis. TOP2A was the most sensitive (97%) biomarker for the detection of HSIL and CDKN2A/p16 the most specific (78%). The combination of TOP2A and CDKN2A/p16 showed a sensitivity of 96% (95% confidence interval (CI): 88-99) and a specificity of 71% (95% CI: 55-82). In the immunohistochemistry analysis, all biomarkers showed a high sensitivity but low specificity for HSIL, except CDKN2A/p16 which had a sensitivity of 100% and a specificity of 63%. The combination of TOP2A and CDKN2A/p16 showed a sensitivity of 100% (95% CI: 91-100) and a specificity of 43% (95% CI: 32-55). The detection of mRNA of cell biomarkers in liquid-based cytology material is feasible. The combination TOP2A and CDKN2A/p16 has a good balance between sensitivity and specificity for the detection of women with HSIL.
authors.affiliation: Faculty of Medicine, Institut Clinic of Gynecology, Obstetrics and Neonatology, Hospital Clínic, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Del Pino
authors.affiliation: Cepheid, Solna, Sweden.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Svanholm-Barrie
authors.affiliation: Faculty of Medicine, Institut Clinic of Gynecology, Obstetrics and Neonatology, Hospital Clínic, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), University of Barcelona, Barcelona, Spain.
authors.forename: Aureli
authors.initials: A
authors.lastname: Torné
authors.affiliation: Faculty of Medicine, Department of Pathology, CRESIB (Centre de Recerca en Salut Internacional de Barcelona), Hospital Clínic, University of Barcelona, Barcelona, Spain.
authors.forename: Lorena
authors.initials: L
authors.lastname: Marimon
authors.affiliation: Cepheid, Solna, Sweden.
authors.forename: Jina
authors.initials: J
authors.lastname: Gaber
authors.affiliation: Faculty of Medicine, Department of Pathology, CRESIB (Centre de Recerca en Salut Internacional de Barcelona), Hospital Clínic, University of Barcelona, Barcelona, Spain.
authors.forename: Amaia
authors.initials: A
authors.lastname: Sagasta
authors.affiliation: Cepheid, Sunnyvale, CA, USA.
authors.forename: David H
authors.initials: DH
authors.lastname: Persing
authors.affiliation: Faculty of Medicine, Department of Pathology, CRESIB (Centre de Recerca en Salut Internacional de Barcelona), Hospital Clínic, University of Barcelona, Barcelona, Spain.
authors.forename: Jaume
authors.initials: J
authors.lastname: Ordi
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis / Q000096:biosynthesis; D014408:Biomarkers, Tumor / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000032:analysis / Q000096:biosynthesis; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000032:analysis / Q000096:biosynthesis; D003581:Cytodiagnosis / Q000379:methods; D004250:DNA Topoisomerases, Type II / Q000032:analysis / Q000096:biosynthesis; D004279:DNA, Viral / Q000032:analysis; D004268:DNA-Binding Proteins / Q000032:analysis / Q000096:biosynthesis; D055088:Early Detection of Cancer / Q000379:methods*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D051034:Inhibitor of Apoptosis Proteins / Q000032:analysis / Q000096:biosynthesis; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis / Q000096:biosynthesis; D020780:Matrix Metalloproteinase 9 / Q000032:analysis / Q000096:biosynthesis; D009687:Nuclear Proteins / Q000032:analysis / Q000096:biosynthesis; D000075223:Poly-ADP-Ribose Binding Proteins; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D000077022:Survivin; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000517:prevention & control; D014626:Vaginal Smears; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis* / Q000517:prevention & control
chemical_list: D000951:Antigens, Neoplasm; C107489:BIRC5 protein, human; D014408:Biomarkers, Tumor; D018797:Cell Cycle Proteins; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D051034:Inhibitor of Apoptosis Proteins; D047908:Intracellular Signaling Peptides and Proteins; C489423:MCM5 protein, human; C492406:NIFK protein, human; D009687:Nuclear Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D012333:RNA, Messenger; D000077022:Survivin; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9; D004250:DNA Topoisomerases, Type II; C000618811:TOP2A protein, human