表达单纯疱疹病毒胸苷激酶的溶瘤腺病毒靶向癌症治疗:在体外评估  

An Oncolytic Adenovirus Expressing Herpes Simplex Virus-thymidine Kinase for Targeting Cancer Therapy: An in vitro Evaluation

作　　者：郑斐群[1] 许尹[2] 吴彬[2] 

机构地区：[1]深圳市第三人民医院放射科,广东深圳518020 [2]中国人民解放军军事医学科学院放射与辐射研究所血液病实验室,北京100850

出　　处：《中国医药指南》2014年第22期84-87,共4页Guide of China Medicine

摘　　要：目的我们希望评估与单纯疱疹病毒胸苷激酶相结合的溶瘤腺病毒治疗效果是否更好。方法我们构建了一种新型的溶瘤腺病毒Ad-ETK,它包含人端粒酶逆转录酶(hTERT)启动子,E1A基因的编码序列,内部核糖体进入位点序列(IRES)和疱疹单纯病毒胸苷激酶(HSV-TK)编码序列。腺病毒Ad-ETK感染了各种肿瘤细胞,并通过用RT-PCR表明的E1A和HSV-TK基因的表达。同时测定了病毒的溶瘤能力及其与HSV-TK系统协同效果。并采用流式细胞仪测量病毒感染效率。结果溶瘤腺病毒Ad-ETK表达了E1A和HSV-TK基因,并且选择性的hTERT-阳性的肿瘤细胞中复制,复制的子代病毒可达150 IU/cell。在体外研究表明,Ad-ETK加更昔洛韦(GCV)有明显的导致细胞死亡的效果。结论溶瘤腺病毒加HSV-TK/GCV自杀基因显著改善治疗效果,在活体评估的结果预示的溶瘤病毒有很好的开发前景。Objective We wish to evaluate whether a strategy that combines the herpes simplex virus-thym idine kinase with oncolytic effects offers a therapeutic advantage.Methods A novel adenovirus Ad-ETK containing a sequentially positioned promoter of human telomerase reverse transcriptase（hTERT）, the coding sequence of E1A gene, an internal ribosome entry site sequence（IRES）and the coding sequence of herpes simplex virus. thymidine kinase（HSV-TK）was constructed.Infection of various cells with Ad-ETK followed by RT-PCR confirmed the expression of EIA and HSV-TK.The oncolytic ability and synergism between oncolytic effects and HSV-TK system was measured.The infection eficiency was determined by flow cytometry. Results Ad-ETK deliverys E1A and HSV-TK gene, which selectively replicates in hTERT-positive tumor cells, and the progeny virus can reach up to 150 IU/cel1.Our in vitro study showed that Ad-ETK plus ganciclovir（GCV） induced an obvious cell death.Conclusion An oncolytic adenovirus plus the HSV-TK/GCV suicide gene system resulted in a significant improvement in treatment efficacy and it may offer important considerations in the development and preclinical assessments of oncolytic virotherapy.

关 键 词：条件复制腺病毒 肿瘤基因治疗 单纯疱疹病毒胸苷激酶 

分 类 号：R752.11[医药卫生—皮肤病学与性病学]