Description
Representing the most relevant procedures and technologies aiding the advance of the field of HPV-mediated carcinogenesis of the cervix and other anatomical regions of squamocolumnar transition, such as the anorectum, penis, and oropharynx, Cervical Cancer: Methods and Protocols compiles a detailed collection of practical chapters. The first half of the book covers HPV types, pathogenesis of cervical cancer (CxCA), prevention, and novel potential drug targets, while the second half explores pathology, genomics, modeling of CxCA, and experimental therapeutic strategies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and vital, Cervical Cancer: Methods and Protocols serves as a valuable resource to both bench scientists and clinicians who step into the realm of high-risk HPVs and CxCA for the first time or those who wish to learn novel approaches or expand their toolbox for the study of CxCA. Part I: Approaches to the Detection and Analysis of HPV Types 1. Evolution and Classification of Oncogenic Human Papillomavirus Types and Variants Associated with Cervical Cancer Zigui Chen, Luciana Bueno de Freitas, and Robert D. Burk 2. A Real-Time PCR Approach Based on SPF10 Primers and the INNO-LiPA HPV Genotyping Extra Assay for the Detection and Typing of Human Papillomavirus M. Isabel Micalessi, Gaelle A. Boulet, and J. Jean-Paul Bogers Part II: Molecular Pathogenesis of CxCA 3. Replication of Human Papillomavirus in Culture Eric J. Ryndock, Jennifer Biryukov, and Craig Meyers 4. HPV Binding Assay to Laminin-332/Integrin a64 on Human Keratinocytes Sarah A. Brendle and Neil D. Christensen 5. Methods to Assess the Nucleocytoplasmic Shuttling of the HPV E1 Helicase and Its Effects on Cellular Proliferation and Induction of a DNA Damage Response Michal Lehoux, Amlie Fradet-Turcotte, and Jacques Archambault 6. Genetic Methods for Studying the Role of Viral Oncogenes in the HPV Life Cycle Jason M. Bodily 7. Robust HPV-18 Production in Organotypic Cultures of Primary Human Keratinocytes Hsu-Kun Wang, Thomas R. Broker, and Louise T. Chow 8. A High-Throughput Cellular Assay to Quantify the p53-Degradation Activity of E6 from Different Human Papillomavirus Types David Gagnon and Jacques Archambault 9. Retroviral Expression of Human Cystatin Genes in HeLa Cells Crystal M. Diep, Gagandeep Kaur, Daniel Keppler, and Athena W. Lin Part III: Vaccinations and Immune Responses to HPV Infections 10. Molecular Analysis of Human Papillomavirus Virus-Like Particle Activated Langerhans Cells In Vitro Andrew W. Woodham, Adam B. Raff, Diane M. Da Silva, and W. Martin Kast Part IV: Identification of Novel Potential Drug Targets 11. Selective Silencing of Gene Target Expression By siRNA Expression Plasmids in Human Cervical Cancer Cells Peralta-Zaragoza Oscar, De la O-Gmez Faustino, Deas Jessica, Fernndez-Tilapa Gloria, Fierros-Zrate Geny del Socorro, Gmez-Cern Claudia, Burguete-Garca Ana, Torres-Poveda Kirvis, Bermdez-Morales Victor Hugo, Rodrguez-Dorantes Mauricio, Prez-Plasencia Carlos, and Madrid-Marina Vicente 12. Silencing of E6/E7 Expression in Cervical Cancer Stem-Like Cells Wenyi Gu, Nigel McMillan, and Chengzhong Yu 13. Two-Step Procedure for Evaluating Experimentally Induced DNA Damage: Texas Red and Comet Assays Gina J. Ferris, Lauren B. Shunkwiler, and Charles A. Kunos 14. Measurement of Deubiquitinating Enzyme Activity Via a Suicidal HA-Ub-VS Probe Colleen Rivard and Martina Bazzaro Part V: Approaches to Detecting, Typing, and Staging of CxCA 15. Immunocytochemical Analysis of the Cervical Pap Smear Terry K. Morgan and Michelle Berlin 16. Diagnosis of HPV-Negative, Gastric-Type Adenocarcinoma of the Endocervix Edyta C. Pirog 17. Targeting of the HPV-16 E7 Protein by RNA Aptamers Julia Dolores Toscano-Garibay, Mara Luisa Bentez-Hess, and Luis Marat Alvarez-Salas 18. The Use of MYBL2 as a Novel Candidate Biomarker of Cervical Cancer Cara M. Martin, Katharine Astbury, Louise Kehoe, Jacqueline Barry O’Crowley, Sharon O’Toole, and John J. O’Leary 19. Fixation Methods for the Preservation of Morphology, RNAs, and Proteins in Paraffin-Embedded Human Cervical Cancer Cell Xenografts in Mice Yoko Matsuda and Toshiyuki Ishiwata Part VI: Genomic Analyses of CxCA 20. Assessment of the HPV DNA Methylation Status in Cervical Lesions Mina Kalantari and Hans-Ulrich Bernard 21. MeDIP-on-Chip for Methylation Profiling Yaw-Wen Hsu, Rui-Lan Huang, and Hung-Cheng Lai 22. Use of DBD-FISH for the Study of Cervical Cancer Progression Eva I. Corts-Gutirrez, Jose L. Fernndez, Martha I. Dvila-Rodrguez, Carmen Lpez-Fernndez, and Jaime Goslvez Part VII: Cellular Assays and Cell Culture Models of CxCA 23. A Quantitative and High-Throughput Assay of Human Papillomavirus DNA Replication David Gagnon, Amlie Fradet-Turcotte, and Jacques Archambault 24. Native Human Papillomavirus Production, Quantification, and Infectivity Analysis Jennifer Biryukov, Linda Cruz, Eric J. Ryndock, and Craig Meyers 25. Functional Analysis of HPV-Like Particle-Activated Langerhans Cells In Vitro Lisa Yan, Andrew W. Woodham, Diane M. Da Silva, and W. Martin Kast 26. Assessment of the Radiation Sensitivity of Cervical Cancer Cell Lines Mayil Vahanan Bose and Thangarajan Rajkumar Part VIII: Animal Models of CxCA and Experimental Therapeutic Strategies 27. Mouse Model of Cervicovaginal Papillomavirus Infection Nicolas uburu, Rebecca J. Cerio, Cynthia D. Thompson, and Patricia M. Day 28. Establishment of Orthotopic Primary Cervix Cancer Xenografts Naz Chaudary, Karolina Jaluba, Melania Pintilie, and Richard P. Hill 29. Generation of K14-E7/N87cat Double Transgenic Mice as a Model of Cervical Cancer Glay Blt and Aykut ren
