title: Increased cycling cell numbers and stem cell associated proteins as potential biomarkers for high grade human papillomavirus+ve pre-neoplastic cervical disease.
abstract: High risk (oncogenic) human papillomavirus (HPV) infection causes cervical cancer. Infections are common but most clear naturally. Persistent infection can progress to cancer. Pre-neoplastic disease (cervical intraepithelial neoplasia/CIN) is classified by histology (CIN1-3) according to severity. Cervical abnormalities are screened for by cytology and/or detection of high risk HPV but both methods are imperfect for prediction of which women need treatment. There is a need to understand the host virus interactions that lead to different disease outcomes and to develop biomarker tests for accurate triage of infected women. As cancer is increasingly presumed to develop from proliferative, tumour initiating, cancer stem cells (CSCs), and as other oncogenic viruses induce stem cell associated gene expression, we evaluated whether presence of mRNA (detected by qRT-PCR) or proteins (detected by flow cytometry and antibody based proteomic microarray) from stem cell associated genes and/or increased cell proliferation (detected by flow cytometry) could be detected in well-characterised, routinely collected cervical samples from high risk HPV+ve women. Both cytology and histology results were available for most samples with moderate to high grade abnormality. We found that stem cell associated proteins including human chorionic gonadotropin, the oncogene TP63 and the transcription factor SOX2 were upregulated in samples from women with CIN3 and that the stem cell related, cell surface, protein podocalyxin was detectable on cells in samples from a subset of women with CIN3. SOX2, TP63 and human gonadotrophin mRNAs were upregulated in high grade disease. Immunohistochemistry showed that SOX2 and TP63 proteins clearly delineated tumour cells in invasive squamous cervical cancer. Samples from women with CIN3 showed increased proliferating cells. We believe that these markers may be of use to develop triage tests for women with high grade cervical abnormality to distinguish those who may progress to cancer from those who may be treated more conservatively.
authors.affiliation: Human Papillomavirus Group, University of Edinburgh, Edinburgh, United Kingdom; Medical Research Council Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Maurice
authors.initials: M
authors.lastname: Canham
authors.affiliation: Medical Research Council Centre for Inflammation Research, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Chara
authors.initials: C
authors.lastname: Charsou
authors.affiliation: Medical Research Council Centre for Inflammation Research, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: June
authors.initials: J
authors.lastname: Stewart
authors.affiliation: Medical Research Council Centre for Inflammation Research, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Sharon
authors.initials: S
authors.lastname: Moncur
authors.affiliation: Medical Research Council Centre for Inflammation Research, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Laura
authors.initials: L
authors.lastname: Hoodless
authors.affiliation: Human Papillomavirus Group, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Ramya
authors.initials: R
authors.lastname: Bhatia
authors.affiliation: Medical Research Council Centre for Inflammation Research, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Duanduan
authors.initials: D
authors.lastname: Cong
authors.affiliation: Human Papillomavirus Group, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Heather
authors.initials: H
authors.lastname: Cubie
authors.affiliation: Simpson Centre for Reproductive Health, Royal Infirmary of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Camille
authors.initials: C
authors.lastname: Busby-Earle
authors.affiliation: Simpson Centre for Reproductive Health, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Alistair
authors.initials: A
authors.lastname: Williams
authors.affiliation: Scottish National Blood Transfusion Service National Science Laboratory, Edinburgh, United Kingdom.
authors.forename: Victoria
authors.initials: V
authors.lastname: McLoughlin
authors.affiliation: Medical Research Council Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, United Kingdom; Scottish National Blood Transfusion Service National Science Laboratory, Edinburgh, United Kingdom.
authors.forename: John D M
authors.initials: JD
authors.lastname: Campbell
authors.affiliation: Scottish Human Papillomavirus Reference Laboratory, Royal Infirmary of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Kate
authors.initials: K
authors.lastname: Cuschieri
authors.affiliation: Medical Research Council Centre for Inflammation Research, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Sarah
authors.initials: S
authors.lastname: Howie
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D005838:Genotype; D052681:Gonadotrophs / Q000378:metabolism; D006801:Humans; D006965:Hyperplasia / Q000473:pathology*; D007150:Immunohistochemistry; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000378:metabolism; D027383:Papillomaviridae / Q000235:genetics; D016133:Polymerase Chain Reaction; D040901:Proteomics; D012333:RNA, Messenger / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D012720:Severity of Illness Index; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation; D002577:Uterine Cervical Diseases / Q000175:diagnosis* / Q000473:pathology / Q000821:virology
chemical_list: D015415:Biomarkers; D004279:DNA, Viral; D012333:RNA, Messenger; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; C115051:TP63 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins