腺病毒载体介导的HSV-tk基因杀伤肿瘤细胞的研究  

Study on Adenovirus Vector Mediated HSV-tk Gene Killing Effect on Tumor Cells

作　　者：李凡[1] 陈立[1] 黄红兰[1] 易世红[1] 赵春燕 魏强[1] 

机构地区：[1]白求恩医科大学病原生物学教研室,长春130021 [2]长春市卫生防疫站

出　　处：《中国生物制品学杂志》2001年第1期4-6,共3页Chinese Journal of Biologicals

摘　　要：目的 探讨ＨＳＶ－ｔｋ基因对各种肿瘤细胞的杀伤作用及人、鼠瘤细胞对其敏感性及瘤细胞表面的αｖ 整合素对腺病毒载体转染的影响。方法用同源重组法构建 ＡｄＣＭＶ ｔｋ基因；蚀斑形成法对重组腺病毒进行扩增、 滴定；Ｘ－ｇａｌ染色法观察腺病毒转染率，结晶紫染色法检测 ＡｄＣＭＶｔｋ／ＧＣＶ的杀伤作用。结果 ＡｄＣＭＶｔｋ对肿瘤细胞 的杀伤强度在一定范围内与ＡｄＣＭＶｔｋ的剂量呈正相关；对人类的Ｈｅｌａ、ＧＲＣ、Ｈｅｐ－２瘤细胞达１００％转染所需的腺病 毒感染复数量（ｍｕｌｔｉｐｌｉｃｉｔｙ　ｏｆ　ｉｎｆｅｃｔｉｏｎ，ｍ．ｏ．ｉ）较鼠Ｌｅｗｉｓ、ＢＴＴ７３９瘤细胞的ＭＯＩ量低；在相同ＭＯＩ病毒量时，有αｖ整合 素的瘤细胞转染率高于无αｖ整合素的瘤细胞。结论ＡｄＣＭＶｔｋ／ＧＣＶ具有较明显的杀伤肿瘤细胞的作用，且有种属 差异，对人瘤细胞作用大于鼠瘤细胞，瘤细胞表面的αｖ整合素有利于腺病毒的转染。Objective To study on the killing effect of HSV-tk gene on tumor cells.Methods The recombinant replication-defective adenovirus of HSV-tk(AdCMVtk)was constructed by homologous recombination techniques,then amplified and titrated by plaque formation test.The transfection efficiency of the virus was assessed by X-gal staining.The killing effect of AdCMVth/CGV was determined by measuring the number of survival cells with crystal violet staining.Results Within certain limits,the killing effect of AdCMVtk on tumor cells was positively related to the dose of it.Murine tumor cells(Lews,BBT739)needed more adenovirus for 100% transfection than human tumor cells(Hela,GRC,Hep-2).The transfection effciency of adenovirus in tumor cells with intergrinαv was higher than those without intergrin αv.Conclusion AdCMVtk/GCV showed significant killing effect on tumor cells.Howerer,the effects were different in various genus-species.The transfection of adenovirus was related to the intergrin αv of tumor cells.

关 键 词：腺病毒 HSK-tk 转染 肿瘤细胞 

分 类 号：R373[医药卫生—病原生物学] R730.51[医药卫生—基础医学]