摘要
背景与目的:以腺病毒为载体的单纯疱疹病毒胸苷激酶基因(adenovirusvector-mediatedherpessimplexvirus-thymidinekinasegene,ADV-TK)重组体是肿瘤基因治疗应用的主要方法之一,本文旨在鉴定自行构建的ADV-TK重组体的体外抗肿瘤活性。方法:利用腺病毒载体将TK导入体外培养的14种不同组织源性的肿瘤细胞,再加入更昔洛韦(ganciclovir,GCV),用MTT法观察对肿瘤细胞的抑制效应。结果:ADV-TK剂量为每孔1×109病毒颗粒,在100μg/mlGCV底物浓度条件下,它对14种肿瘤细胞中的11种杀伤率达74%以上,而对人喉上皮癌细胞Hep-2、人肝癌细胞Bel-7402和人结肠癌细胞HCT-8敏感性稍差,分别为(55.3±2.0)%、(61.3±2.0)%和(63.7±2.5)%,ADV-TK对肿瘤细胞的抑制率除人喉上皮癌细胞Hep-2外,与相当于组织峰值药物浓度(5μg/ml)的化疗药物顺铂的效率相近。结论:ADV-TK重组体具有明确的体外抗肿瘤活性,有潜在临床应用价值。
BACKGROUND &OBJECTIVE: Adenovirus vector-mediated herpes simpl ex virus thymidine kinase gene (ADV-TK) transfer is one of the major gene therapy strategies for tumor. This study was designed to determine the in vitro anti-t umor efficiency of ADV-TK, a recombinant construct previously developed in our laboratory. METHODS: Herpes simplex virus thymidine kinase (TK) gene was transdu ced into 14 types of cultured tumor cells with different histological origins us ing adenovirus vector followed by ganciclovir (GCV) medication. The killing effi ciency was measured using MTT assay. RESULTS: At the dosage of 1×109 viral part icles/per well in the presence of 100 μg/ml GCV, ADV-TK/GCV caused effective k illing of 11 out of total 14 types of tumor cells with a rate higher than 74%, the other 3 tumor cells, laryngeal epithelial cancer cells (Hep-2), hepatic can cer cells (Bel7402), and human colon cancer cells (HCT-8) were less sensitive t o the ADV-TK/GCV treatment with the killing rates of 55.3%±2.0%, 61.3%±2.0 %, and 63.7%±2.5%, respectively. Except for Hep-2, the killing efficiency c aused by ADV-TK/GCV treatment was similar to that caused by cisplatin at a dosa ge equal to the in vivo peak concentration (5 μg/ml) in tissue. CONCLUSION: ADV -TK is highly efficient for active killing of tumor cells in vitro and is promi sing for future clinical application.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2003年第12期1264-1267,共4页
Chinese Journal of Cancer
基金
国家重大科技专项"创新药物
中药现代化""863"项目(No.2002AA2Z3348)
关键词
重组腺病毒
胸苷激酶
基因
肿瘤细胞
杀伤作用
Adenovirus vector
Herpes simplex virus thymidine kinase gene
Gene therapy
Tumor cell
