title: Artificial microRNAs against the viral E6 protein provoke apoptosis in HPV positive cancer cells.
abstract: High-risk human papillomavirus (HPV) types 16 and 18 are associated with more than 70% of cervical cancer cases. The oncoprotein E6 is multifunctional and has numerous cellular partners. The best-known activity of E6 is the polyubiquination of the pro-apoptotic tumor suppressor p53, targeting it for degradation by the 26S proteasome. Loss of p53 triggers genomic instability and favors cancer development. Here, we generated recombinant adenovirus (Ad) vectors expressing artificial microRNAs directed against HPV16 E6 (Ad16_1) or HPV18 E6 (Ad18_2). E6-knockdown was observed in HeLa after treatment with Ad18_2 and in SiHa with Ad16_1. Western-blot experiments found an increase in p53 levels after treatment in both cell lines. Cell death was observed in both cell lines after knockdown of E6. Further analysis such as cleavage of caspases (3 and 7) as well as of PARP1 indicated that treated HeLa and SiHa cells underwent apoptosis. The growth of HeLa-derived tumors developed in nude mice was significantly reduced after intra-tumoral injection of Ad18_2. Therefore, vectorisation of artificial miRNA against E6 oncoprotein by means of recombinant adenoviruses might represent a valuable therapeutic approach for treating HPV-positive cancers.
authors.affiliation: UMR7242, IREBS, Illkirch, Université de Strasbourg, CNRS, France.
authors.forename: Anaëlle Charlotte
authors.initials: AC
authors.lastname: Bonetta
authors.affiliation: INSERM, Unité 1110, Institut de Recherche sur les Maladies Virales et Hépatiques, Strasbourg, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Mailly
authors.affiliation: INSERM, Unité 1110, Institut de Recherche sur les Maladies Virales et Hépatiques, Strasbourg, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Robinet
authors.affiliation: UMR7242, IREBS, Illkirch, Université de Strasbourg, CNRS, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Travé
authors.affiliation: UMR7242, IREBS, Illkirch, Université de Strasbourg, CNRS, France.
authors.forename: Murielle
authors.initials: M
authors.lastname: Masson
authors.affiliation: UMR7242, IREBS, Illkirch, Université de Strasbourg, CNRS, France. Electronic address: francois.deryckere@unistra.fr.
authors.forename: François
authors.initials: F
authors.lastname: Deryckere
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D005260:Female; D055785:Gene Knockdown Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006367:HeLa Cells; D052162:Human papillomavirus 16 / Q000235:genetics / Q000472:pathogenicity; D052161:Human papillomavirus 18 / Q000235:genetics / Q000472:pathogenicity; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics*; D009856:Oncogene Proteins, Viral / Q000037:antagonists & inhibitors* / Q000235:genetics*; D030361:Papillomavirus Infections / Q000473:pathology / Q000628:therapy* / Q000821:virology; D012097:Repressor Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D002583:Uterine Cervical Neoplasms / Q000473:pathology / Q000628:therapy* / Q000821:virology; D023041:Xenograft Model Antitumor Assays
chemical_list: D004268:DNA-Binding Proteins; C068651:E6 protein, Human papillomavirus type 16; C052603:E6 protein, Human papillomavirus type 18; D035683:MicroRNAs; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53
keywords: Adenovirus; Cancer therapy; Cervical cancer; Gene therapy; RNA interference