title: Different DNA damage and cell cycle checkpoint control in low- and high-risk human papillomavirus infections of the vulva.
abstract: Human papillomavirus (HPV) infections may result in benign hyperplasia, caused by low-risk HPV types, or (pre)malignant lesions caused by high-risk HPV types. The molecular basis of this difference in malignant potential is not completely understood. Here, we performed gene profiling of different HPV infected vulvar tissues (condylomata acuminata (n = 5), usual type vulvar intraepithelial neoplasia (uVIN) (n = 9)) and control samples (n = 14) using Affymetrix Human U133A plus 2 GeneChips. Data were analyzed using OmniViz®, Partek® and Ingenuity® Software. Results were validated by real-time RT-PCR and immunostaining. Although similarities were observed between gene expression profiles of low- and high-risk HPV infected tissues (e.g., absence of estrogen receptor in condylomata and uVIN), high-risk HPV infected tissues showed more proliferation and displayed more DNA damage than tissues infected with low-risk HPV. These observations were confirmed by differential regulation of cell cycle checkpoints and by increased expression of DNA damage-biomarkers p53 and γH2AX. Furthermore, FANCA, FANCD2, BRCA1 and RAD51, key players in the DNA damage response, were significantly upregulated (p < 0.05). In addition, we compared our results with publicly available gene expression profiles of various other HPV-induced cancers (vulva, cervix and head-and-neck). This showed p16(INK4a)  was the most significant marker to detect a high-risk HPV infection, but no other markers could be found. In conclusion, this study provides insight into the molecular basis of low- and high-risk HPV infections and indicates two main pathways (cell cycle and DNA damage response) that are much stronger affected by high-risk HPV as compared to low-risk HPV.
authors.affiliation: Department of Obstetrics and Gynaecology, Erasmus Medical Center, Rotterdam, The Netherlands. l.santegoets@erasmusmc.nl
authors.forename: Lindy A M
authors.initials: LA
authors.lastname: Santegoets
authors.forename: Romy
authors.initials: R
authors.lastname: van Baars
authors.forename: Annelinde
authors.initials: A
authors.lastname: Terlou
authors.forename: Claudia
authors.initials: C
authors.lastname: Heijmans-Antonissen
authors.forename: Sigrid M A
authors.initials: SM
authors.lastname: Swagemakers
authors.forename: Peter J
authors.initials: PJ
authors.lastname: van der Spek
authors.forename: Patricia C
authors.initials: PC
authors.lastname: Ewing
authors.forename: Marc
authors.initials: M
authors.lastname: van Beurden
authors.forename: Willem I
authors.initials: WI
authors.lastname: van der Meijden
authors.forename: Theo J M
authors.initials: TJ
authors.lastname: Helmerhorst
authors.forename: Leen J
authors.initials: LJ
authors.lastname: Blok
mesh_terms: D052685:Alphapapillomavirus; D019313:BRCA1 Protein / Q000096:biosynthesis; D014408:Biomarkers, Tumor; D059447:Cell Cycle Checkpoints; D003218:Condylomata Acuminata / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000821:virology; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D052217:Fanconi Anemia Complementation Group A Protein / Q000096:biosynthesis; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000096:biosynthesis; D005260:Female; D020869:Gene Expression Profiling; D006657:Histones / Q000096:biosynthesis; D006801:Humans; D030361:Papillomavirus Infections / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000821:virology; D051135:Rad51 Recombinase / Q000096:biosynthesis; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis; D014844:Vulva / Q000473:pathology* / Q000821:virology; D014845:Vulvar Diseases / Q000235:genetics* / Q000473:pathology / Q000821:virology
chemical_list: D019313:BRCA1 Protein; C492913:BRCA1 protein, human; D014408:Biomarkers, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16; D004279:DNA, Viral; C498471:FANCA protein, human; C499905:FANCD2 protein, human; D052217:Fanconi Anemia Complementation Group A Protein; D052236:Fanconi Anemia Complementation Group D2 Protein; C468783:H2AX protein, human; D006657:Histones; D016159:Tumor Suppressor Protein p53; C495153:RAD51 protein, human; D051135:Rad51 Recombinase