Kari Ilkka, Syrjänen Stina, Johansson Bo, Peri Piritta, He Bin, Roizman Bernard, Hukkanen Veijo
Department of Virology, Institute of Dentistry, University of Turku, Turku, Finland.
Virol J. 2007 Jun 4;4:47. doi: 10.1186/1743-422X-4-47.
Human papillomavirus (HPV) infection is known to be the most important etiologic factor of cervical cancer. There is no HPV specific therapy available for treatment of invasive squamous cell carcinoma of the cervix and its precursor lesions. The present study elucidates the potential to use herpes simplex virus (HSV) derived vectors for expression of antisense RNA to HPV -16 E7 oncogene.
We have constructed replication competent, nonneuroinvasive HSV-1 vectors, deleted of the gamma134.5 gene. The vectors express RNA antisense to the first 100 nucleotides of the HPV-16 E7 gene. We assayed the ability of the antisense E7 vectors R5225 (tk-) and R5226 (tk+), to produce antisense RNA, as well as the consequent effects on E7 mRNA and protein levels in HPV-16 positive CaSki cells. Anti-E7 RNA was expressed by both constructs in a dose-dependent manner. Expression of HPV-16 E7 mRNA was downregulated effectively in CaSki cells infected with the tk- recombinant R5225 or with R5226. The tk+ recombinant R5226 was effective in downregulating E7 protein expression.
We have shown that anti-E7 RNA expressed from an HSV vector could efficiently downregulate HPV-16 E7 mRNA and E7 protein expression in CaSki cells. We conclude that HSV vectors may become a useful tool for gene therapy of HPV infections.
人乳头瘤病毒（HPV）感染是已知的宫颈癌最重要的病因。目前尚无针对宫颈浸润性鳞状细胞癌及其前驱病变的HPV特异性治疗方法。本研究阐明了使用单纯疱疹病毒（HSV）衍生载体表达针对HPV -16 E7癌基因的反义RNA的潜力。
我们构建了具有复制能力、非神经侵袭性的HSV-1载体，缺失了γ134.5基因。这些载体表达针对HPV-16 E7基因前100个核苷酸的反义RNA。我们检测了反义E7载体R5225（tk-）和R5226（tk+）产生反义RNA的能力，以及对HPV-16阳性CaSki细胞中E7 mRNA和蛋白质水平的后续影响。两种构建体均以剂量依赖方式表达抗E7 RNA。在感染tk-重组体R5225或R5226的CaSki细胞中，HPV-16 E7 mRNA的表达被有效下调。tk+重组体R5226在下调E7蛋白表达方面有效。
我们已经表明，从HSV载体表达的抗E7 RNA可以有效下调CaSki细胞中HPV-16 E7 mRNA和E7蛋白的表达。我们得出结论，HSV载体可能成为HPV感染基因治疗的有用工具。
