title: Strong inverse correlation between microRNA-125b and human papillomavirus DNA in productive infection.
abstract: Infection by the human papillomavirus (HPV) is a cause of cervical intraepithelial neoplasia (CIN) and cancer. microRNA (miRNA) in situ analysis of the transformation zone epithelia, the site of initial cervical HPV infection, showed that miRNAs let-7c, -99a, 26a, and 125b were the most abundantly expressed. In situ testing of CIN 1 showed a dramatic reduction in miR-125b expression in the koilocytes, the cytologic marker of productive HPV infection. A marked reduction in miR-125b was likewise observed in the HPV-infected cells of the condyloma acuminatum, verruca vulgaris, and epidermodysplasia verruciformis. Reverse transcriptase in situ polymerase chain reaction (PCR) showed that the pre-miRNA 125b was present in the koilocyte, suggesting direct inactivation of the mature miRNA. HEK cells transfected with only the antimiR-125b showed perinuclear halos equivalent to HPV-infected koilocytes. NIH 3T3 cells transfected with the HPV 16 full-length genome and mimetic miR-125b showed a marked reduction in viral DNA and protein synthesis by quantitative PCR and in situ-based analyses, respectively (P=0.002). Alternatively, cotransfection with anti-miR-125b and HPV 16 markedly increased HPV DNA (P=0.002). Sequence analyses showed strong homology between L2 of different HPV genotypes and miR-125b. Transfection with HPV 16 L2 resulted in a marked reduction in miR-125b levels in the NIH 3T3 cells. HPV L2-induced inactivation of miR-125b is associated with the classic cytologic changes of the koilocyte, and the exogenous application of mimetic miR-125b markedly inhibits HPV DNA synthesis.
authors.affiliation: The Comprehensive Cancer Center, Rio de Janeiro, Brazil. gnuovomd@pol.net
authors.forename: Gerard J
authors.initials: GJ
authors.lastname: Nuovo
authors.forename: Xin
authors.initials: X
authors.lastname: Wu
authors.forename: Stefano
authors.initials: S
authors.lastname: Volinia
authors.forename: Fengting
authors.initials: F
authors.lastname: Yan
authors.forename: Gianpiero
authors.initials: G
authors.lastname: di Leva
authors.forename: Nena
authors.initials: N
authors.lastname: Chin
authors.forename: Alcina F
authors.initials: AF
authors.lastname: Nicol
authors.forename: Jinmai
authors.initials: J
authors.lastname: Jiang
authors.forename: Gregory
authors.initials: G
authors.lastname: Otterson
authors.forename: Thomas D
authors.initials: TD
authors.lastname: Schmittgen
authors.forename: Carlo
authors.initials: C
authors.lastname: Croce
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002584:Cervix Uteri / Q000166:cytology / Q000473:pathology; D003218:Condylomata Acuminata / Q000473:pathology / Q000821:virology; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D004819:Epidermodysplasia Verruciformis / Q000473:pathology / Q000821:virology; D004847:Epithelial Cells / Q000473:pathology; D005260:Female; D006801:Humans; D051379:Mice; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics* / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000473:pathology* / Q000821:virology; D014860:Warts / Q000473:pathology / Q000821:virology; D002578:Uterine Cervical Dysplasia / Q000473:pathology / Q000821:virology
chemical_list: D004279:DNA, Viral; C531888:MIRN125 microRNA, human; D035683:MicroRNAs