Increase in viral load, viral integration, and gain of telomerase genes during uterine cervical carcinogenesis can be simultaneously assessed by the HPV 16/18 MLPA-assay.
Theelen, Wendy; Speel, Ernst*-Jan M; Herfs, Michaelet al.
2010 • In American Journal of Pathology, 177 (4), p. 2022-33
[en] Oncogenic human papillomavirus (HPV) infection is the most important risk factor in cervical carcinogenesis cases; high viral loads, viral integration into the host genome, and gain of the telomerase-related genes, TERT and TERC, are all factors associated with progression to cancer. A recently developed multiparameter HPV 16/18 multiplex ligation-dependent probe amplification (MLPA) assay, which allows the simultaneous assessment of these factors, was applied to a series of 67 normal and (pre)malignant frozen uterine cervical samples, as well as to 91 cytological preparations, to test the ability of the MLPA assay to identify high-risk lesions on the basis of these factors. Validation was performed using quantitative PCR, the PapilloCheck and fluorescence in situ hybridization. Only 5 out of 37 normal tissue samples or low-grade cervical lesions (ie, CIN1 and condyloma) showed either an HPV16 viral load higher than 25 copies per cell, viral integration, and/or gain of one of the telomerase-related genes, whereas for the high-grade cervical lesions, one or more of these risk factors was found in 25 of 30 cases. The HPV MLPA assay showed a sensitivity of 83% and a specificity of 86% in frozen cervical specimens. Furthermore, the feasibility of the MLPA assay was shown for cytological samples, where in 57% of high-grade squamous intraepithelial lesion cases, the high-risk factors were detected using this assay.
Research center :
GIGA‐R - Giga‐Research - ULiège
Disciplines :
Immunology & infectious disease Oncology
Author, co-author :
Theelen, Wendy; Departments of Molecular Cell Biology, > GROW - School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, The Netherlands
Speel, Ernst*-Jan M; Departments of Molecular Cell Biology, > GROW - School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, The Netherlands
Herfs, Michael ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Reijans, Martin
Simons, Guus
Meulemans, Els V
Baldewijns, Marcella M
Ramaekers, Frans C S
Somja, Joan ; Centre Hospitalier Universitaire de Liège - CHU > Anatomie pathologique
Delvenne, Philippe ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Hopman, Anton H N
Language :
English
Title :
Increase in viral load, viral integration, and gain of telomerase genes during uterine cervical carcinogenesis can be simultaneously assessed by the HPV 16/18 MLPA-assay.
Publication date :
2010
Journal title :
American Journal of Pathology
ISSN :
0002-9440
eISSN :
1525-2191
Publisher :
American Society for Investigative Pathology, Bethesda, United States - Maryland
Jordan J, Arbyn M, Martin-Hirsch P, Schenck U, Baldauf JJ, Da Silva D, Anttila A, Nieminen P, Prendiville W: European guidelines for quality assurance in cervical cancer screening: recommendations for clinical management of abnormal cervical cytology, part 1. Cytopathology 2008, 19:342-354
Spitzer M, Apgar BS, Brotzman GL: Management of histologic abnormalities of the cervix. Am Fam Physician 2006, 73:105-112
ACOG Practice Bulletin No. 99: management of abnormal cervical cytology and histology. Obstet Gynecol 2008, 112:1419-1444
Cuzick J, Clavel C, Petry KU, Meijer CJ, Hoyer H, Ratnam S, Szarewski A, Birembaut P, Kulasingam S, Sasieni P, Iftner T: Overview of the European and North American studies on HPV testing in primary cervical cancer screening. Int J Cancer 2006, 119:1095-1101
Rao A, Pather S, Dalrymple C, Mackie A, Deans R, Carter J: The role of HPV testing in patients with possible high-grade cervical cytology. J Obstet Gynaecol Res 2009, 35:503-506
Theelen W, Reijans M, Simons G, Ramaekers FCS, Speel EJM, Hopman AHN: A new multiparameter assay to assess HPV 16/18 viral load and physical status together with gain of telomerase genes in HPV-related cancers. Int J Cancer 2010 126:959-975
Schouten JP, McElgunn CJ, Waaijer R, Zwijnenburg D, Diepvens F, Pals G: Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification. Nucleic Acids Res 2002, 30:e57
Shen Y, Wu BL: Designing a simple multiplex ligation-dependent probe amplification (MLPA) assay for rapid detection of copy number variants in the genome. J Genet Genomics 2009, 36:257-265
Munoz N, Bosch FX, de Sanjose S, Herrero R, Castellsague X, Shah KV, Snijders PJ, Meijer CJ: Epidemiologic classification of human papillomavirus types associated with cervical cancer. N Engl J Med 2003, 348:518-527 (Pubitemid 36159887)
Munoz N, Castellsague X, de Gonzalez AB, Gissmann L: Chapter 1: HPV in the etiology of human cancer. Vaccine 2006, 24 (Suppl 3):1-10
Saunier M, Monnier-Benoit S, Mauny F, Dalstein V, Briolat J, Riethmuller D, Kantelip B, Schwarz E, Mougin C, Pretet JL: Analysis of human papillomavirus type 16 (HPV16) DNA load and physical state for identification of HPV16-infected women with high-grade lesions or cervical carcinoma. J Clin Microbiol 2008, 46:3678-3685
Klaes R, Woerner SM, Ridder R, Wentzensen N, Duerst M, Schneider A, Lotz B, Melsheimer P, von Knebel Doeberitz M: Detection of high-risk cervical intraepithelial neoplasia and cervical cancer by amplification of transcripts derived from integrated papillomavirus oncogenes. Cancer Res 1999, 59:6132-6136
Cheung JL, Cheung TH, Ng CW, Yu MY, Wong MC, Siu SS, Yim SF, Chan PK: Analysis of human papillomavirus type 18 load and integration status from low-grade cervical lesion to invasive cervical cancer. J Clin Microbiol 2009, 47:287-293
Hesselink AT, Berkhof J, Heideman DA, Bulkmans NW, van Tellingen JE, Meijer CJ, Snijders PJ: High-risk human papillomavirus DNA load in a population-based cervical screening cohort in relation to the detection of high-grade cervical intraepithelial neoplasia and cervical cancer. Int J Cancer 2009, 124:381-386
Xi LF, Koutsky LA, Castle PE, Wheeler CM, Galloway DA, Mao C, Ho J, Kiviat NB: Human papillomavirus type 18 DNA load and 2-year cumulative diagnoses of cervical intraepithelial neoplasia grades 2-3. J Natl Cancer Inst 2009, 101:153-161
Pirami L, Giache V, Becciolini A: Analysis of HPV16, 18, 31, and 35 DNA in pre-invasive and invasive lesions of the uterine cervix. J Clin Pathol 1997, 50:600-604
Vinokurova S, Wentzensen N, Kraus I, Klaes R, Driesch C, Melsheimer P, Kisseljov F, Durst M, Schneider A, von Knebel Doeberitz M: Typedependent integration frequency of human papillomavirus genomes in cervical lesions. Cancer Res 2008, 68:307-313
Heselmeyer K, Schrock E, du Manoir S, Blegen H, Shah K, Steinbeck R, Auer G, Ried T: Gain of chromosome 3q defines the transition from severe dysplasia to invasive carcinoma of the uterine cervix. Proc Natl Acad Sci USA 1996, 93:479-484
Hopman AHN, Theelen W, Hommelberg PP, Kamps MA, Herrington CS, Morrison LE, Speel EJM, Smedts F, Ramaekers FCS: Genomic integration of oncogenic HPV and gain of the human telomerase gene TERC at 3q26 are strongly associated events in the progression of uterine cervical dysplasia to invasive cancer. J Pathol 2006, 210:412-419
Huang FY, Kwok YK, Lau ET, Tang MH, Ng TY, Ngan HY: Genetic abnormalities and HPV status in cervical and vulvar squamous cell carcinomas. Cancer Genet Cytogenet 2005, 157:42-48
Matthews CP, Shera KA, McDougall JK: Genomic changes and HPV type in cervical carcinoma. Proc Soc Exp Biol Med 2000, 223: 316-321
van den Brule AJ, Pol R, Fransen-Daalmeijer N, Schouls LM, Meijer CJ, Snijders PJ: GP5+/6+ PCR followed by reverse line blot analysis enables rapid and high-throughput identification of human papillomavirus genotypes. J Clin Microbiol 2002, 40:779-787
Lindh M, Gorander S, Andersson E, Horal P, Mattsby-Balzer I, Ryd W: Real-time taqman PCR targeting 14 human papilloma virus types. J Clin Virol 2007, 40:321-324
Peitsaro P, Johansson B, Syrjanen S: Integrated human papillomavirus type 16 is frequently found in cervical cancer precursors as demonstrated by a novel quantitative real-time PCR technique. J Clin Microbiol 2002, 40:886-891
Lee GM, Thornthwaite JT, Rasch EM: Picogram per cell determination of DNA by flow cytofluorometry. Anal Biochem 1984, 137:221-226
Kulmala SM, Shabalova IP, Petrovitchev N, Syrjanen KJ, Gyllensten UB, Johansson BC, Syrjanen SM: Type-specific persistence of highrisk human papillomavirus infections in the New Independent States of the former Soviet Union Cohort Study. Cancer Epidemiol Biomarkers Prev 2007, 16:17-22
Jones J, Powell NG, Tristram A, Fiander AN, Hibbitts S: Comparison of the PapilloCheck DNA micro-array Human Papillomavirus detection assay with Hybrid Capture II and PCR-enzyme immunoassay using the GP5/6+ primer set. J Clin Virol 2009, 45:100-104
Rebolj M, van Ballegooijen M, Berkers LM, Habbema D: Monitoring a national cancer prevention program: successful changes in cervical cancer screening in the Netherlands. Int J Cancer 2007, 120:806-812
Cricca M, Morselli-Labate AM, Venturoli S, Ambretti S, Gentilomi GA, Gallinella G, Costa S, Musiani M, Zerbini M: Viral DNA load, physical status and E2/E6 ratio as markers to grade HPV16 positive women for high-grade cervical lesions. Gynecol Oncol 2007, 106:549-557
Hudelist G, Manavi M, Pischinger KI, Watkins-Riedel T, Singer CF, Kubista E, Czerwenka KF: Physical state and expression of HPV DNA in benign and dysplastic cervical tissue: different levels of viral integration are correlated with lesion grade. Gynecol Oncol 2004, 92: 873-880
Hopman AHN, Smedts F, Dignef W, Ummelen M, Sonke G, Mravunac M, Vooijs GP, Speel EJM, Ramaekers FCS: Transition of high-grade cervical intraepithelial neoplasia to micro-invasive carcinoma is characterized by integration of HPV 16/18 and numerical chromosome abnormalities. J Pathol 2004, 202:23-33
Roberts I, Ng G, Foster N, Stanley M, Herdman MT, Pett MR, Teschendorff A, Coleman N: Critical evaluation of HPV16 gene copy number quantification by SYBR green PCR. BMC Biotechnol 2008, 8:57
Ruutu MP, Kulmala SM, Peitsaro P, Syrjanen SM: The performance of the HPV16 real-time PCR integration assay. Clin Biochem 2008, 41:423-428
Gallo G, Bibbo M, Bagella L, Zamparelli A, Sanseverino F, Giovagnoli MR, Vecchione A, Giordano A: Study of viral integration of HPV-16 in young patients with LSIL. J Clin Pathol 2003, 56:532-536
Heselmeyer-Haddad K, Janz V, Castle PE, Chaudhri N, White N, Wilber K, Morrison LE, Auer G, Burroughs FH, Sherman ME, Ried T: Detection of genomic amplification of the human telomerase gene (TERC) in cytologic specimens as a genetic test for the diagnosis of cervical dysplasia. Am J Pathol 2003, 163:1405-1416
