Martinez I, Gardiner A S, Board K F, Monzon F A, Edwards R P, Khan S A
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Oncogene. 2008 Apr 17;27(18):2575-82. doi: 10.1038/sj.onc.1210919. Epub 2007 Nov 12.
Human papillomaviruses (HPVs) are involved in the pathogenesis of cancer of the cervix (CaCx). MicroRNA (miRNA) expression analysis using Ambion (Austin, TX, USA) arrays showed that three miRNAs were overexpressed and 24 underexpressed in cervical cell lines containing integrated HPV-16 DNA compared to the normal cervix. Furthermore, nine miRNAs were overexpressed and one underexpressed in integrated HPV-16 cell lines compared to the HPV-negative CaCx cell line C-33A. Based on microarray and/or quantitative real-time PCR and northern blot analyses, microRNA-218 (miR-218) was specifically underexpressed in HPV-positive cell lines, cervical lesions and cancer tissues containing HPV-16 DNA compared to both C-33A and the normal cervix. Expression of the E6 oncogene of high-risk HPV-16, but not that of low-risk HPV-6, reduced miR-218 expression, and conversely, RNA interference of E6/E7 oncogenes in an HPV-16-positive cell line increased miR-218 expression. We also demonstrate that the epithelial cell-specific marker LAMB3 is a target of miR-218. We also show that LAMB3 expression is increased in the presence of the HPV-16 E6 oncogene and this effect is mediated through miR-218. These findings may contribute to a better understanding of the molecular mechanisms involved in cervical carcinogenesis.
人乳头瘤病毒(HPV)与宫颈癌(CaCx)的发病机制有关。使用Ambion(美国得克萨斯州奥斯汀)芯片进行的微小RNA(miRNA)表达分析表明,与正常宫颈相比,含有整合型HPV-16 DNA的宫颈细胞系中有3种miRNA过表达,24种miRNA表达不足。此外,与HPV阴性的CaCx细胞系C-33A相比,整合型HPV-16细胞系中有9种miRNA过表达,1种miRNA表达不足。基于芯片和/或定量实时PCR以及Northern印迹分析,与C-33A和正常宫颈相比,微小RNA-218(miR-218)在HPV阳性细胞系、宫颈病变组织和含有HPV-16 DNA的癌组织中特异性表达不足。高危型HPV-16的E6癌基因的表达,而非低危型HPV-6的E6癌基因的表达,降低了miR-218的表达,相反,在HPV-16阳性细胞系中对E6/E7癌基因进行RNA干扰则增加了miR-218的表达。我们还证明上皮细胞特异性标志物LAMB3是miR-218的一个靶标。我们还表明,在存在HPV-16 E6癌基因的情况下LAMB3的表达增加,并且这种作用是通过miR-218介导的。这些发现可能有助于更好地理解宫颈癌发生过程中涉及的分子机制。