Inhibition of cervical cancer cell growth by human papillomavirus virus–like particles packaged with human papillomavirus oncoprotein short hairpin RNAs

• Published in: Molecular cancer therapeutics Vol. 8; no. 2; pp. 357 - 365
• Main Authors: Bousarghin, Latifa, Touze, Antoine, Gaud, Guillaume, Iochmann, Sophie, Alvarez, Eva, Reverdiau, Pascale, Gaitan, Julien, Jourdan, Marie-Lise, Sizaret, Pierre-Yves, Coursaget, Pierre L
• Format: Journal Article
• Language: English
• Published: United States American Association for Cancer Research 01.02.2009
• Subjects: Animals; Apoptosis; Cell Line, Tumor; Cell Proliferation; Chlorides; E6 and E7 oncoproteins; Female; Gene Expression Regulation, Neoplastic; Gene Silencing; gene transfer; Humans; Mice; Oncogene Proteins, Viral - genetics; Oncogene Proteins, Viral - metabolism; Papillomaviridae - physiology; Papillomavirus; Papillomavirus E7 Proteins; pseudovirions; Repressor Proteins - genetics; Repressor Proteins - metabolism; RNA interference; RNA, Small Interfering - metabolism; Transduction, Genetic; Transfection; Uterine Cervical Neoplasms - genetics; Uterine Cervical Neoplasms - pathology; Uterine Cervical Neoplasms - virology; Virion - physiology; Virus Assembly - physiology; Zinc Compounds
• ISSN: 1535-7163; 1538-8514
• DOI: 10.1158/1535-7163.MCT-08-0626

Overexpression of human papillomavirus (HPV E6 and HPV E7) oncogenes in human cervical cells results in the development of cancer, and E6 and E7 proteins are therefore targets for preventing cervical cancer progression. Here, we describe the silencing of E6 and E7 expression in cervical carcinoma cells by RNA interference. In order to increase the efficacy of the RNA interference, HPV pseudovirions coding for a short hairpin RNA (shRNA) sequence were produced. The results indicated the degradation of E6 and E7 mRNAs when shRNA against E6 or E7 were delivered by pseudovirions in HPV-positive cells (CaSki and TC1 cells). E6 silencing resulted in the accumulation of cellular p53 and reduced cell viability. More significant cell death was observed when E7 expression was suppressed. Silencing E6 and E7 and the consequences for cancer cell growth were also investigated in vivo in mice using the capacity of murine TC1 cells expressing HPV-16 E6 and E7 oncogenes to induce fast-growing tumors. Treatment with lentiviruses and HPV virus-like particle vectors coding for an E7 shRNA sequence both resulted in dramatic inhibition of tumor growth. These results show the ability of pseudovirion-delivered shRNA to produce specific gene suppression and provide an effective means of reducing HPV-positive tumor growth. [Mol Cancer Ther 2009;8(2):357–65]