CAJ | 학술논문

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。
목적：아문희망평고여단순포진병독흉감격매상결합적용류선병독치료효과시부경호。방법아문구건료일충신형적용류선병독Ad-ETK，타포함인단립매역전록매（hTERT）계동자，E1A기인적편마서렬，내부핵당체진입위점서렬（IRES）화포진단순병독흉감격매（HSV-TK）편마서렬。선병독Ad-ETK감염료각충종류세포，병통과용RT-PCR표명적E1A화HSV-TK기인적표체。동시측정료병독적용류능력급기여HSV-TK계통협동효과。병채용류식세포의측량병독감염효솔。결과용류선병독Ad-ETK표체료E1A화HSV-TK기인，병차선택성적hTERT-양성적종류세포중복제，복제적자대병독가체150 IU/cell。재체외연구표명，Ad-ETK가경석락위（GCV）유명현적도치세포사망적효과。결론용류선병독가HSV-TK/GCV자살기인현저개선치료효과，재활체평고적결과예시적용류병독유흔호적개발전경。
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.