HSV-tk基因/GCV系统治疗肿瘤的旁观者效应与缝隙连接关系的研究  

Relatonship between Bystander Effect and Gap Junction in Tumor Treated by HSV - tk Gene/GCV System

作　　者：曹剑峰[1] 汪超英[1] 张涛[1] 王志友[1] 李涛[2] 李玲莉[2] 李承晏[2] 余绍祖[2] 

机构地区：[1]武汉生物制品研究所,武汉430060 [2]武汉大学人民医院,武汉430060

出　　处：《中国生物制品学杂志》2002年第1期21-23,共3页Chinese Journal of Biologicals

摘　　要：目的 探讨单纯疱疹病毒胸苷激酶（HSV－tk）基因／丙氧鸟苷（GCV）系统杀伤肿瘤细胞时旁观者效应与细胞间缝隙连接的关系。方法 用PA317细胞包装STK质粒（逆转录病毒载体介导的HSV-tk基因），形成假逆转录病毒颗粒，转染有缝隙连接的大鼠胶质瘤细胞C6和无缝隙连接的入宫颈癌细胞 Hela，制成 C6 tk+和 Helatk+细胞，将不同比例的C6和 C6 tk+、Hela和 Hela tk+细胞混合，每种比例8孔，培养 24h后 4孔加入含0．5μg/mlGCV的培养基，4孔作对照，6d后用MTT法测量细胞存活率。在两个培养皿中放入周围粘有33mm高滤纸的载玻片，分别接种 C6、C6 tk+细胞。细胞 80%汇合后将载玻片互换，并加入含0．5μg/ml GCV的培养基，6d后观察细胞形态。结果C6细胞组存在旁观者效应，Heal细胞组不存在旁观者效应。培养皿中tk+细胞大部分被杀死,tk-细胞无变化。结论HSV-tk基因/GCV系统杀伤肿瘤细胞时旁观者效应与细胞间缝隙连接有关。Abstract: Objective To evaluate the relationship between bystander effect and gap junction duringkilling tumor cells by HSV - tk gene/GCV system. Methods STK plasmid was packed with PA317 cells andtransformed to the rat glioma cell C6 with gap junction and human cervical carcinoma cell Hala without gapjunction. The prepared C6 tk+ and Hela tk + cells were mixed at different ratios with C6 and Hela cells re-spectively, then inoculated to a 96 - well plate (8 wells per ration) and cultured for 24h. The media contain-ing 0.5/ig/ml GCV were added in 4 wells, and survival cells were detected by MTT method 6d later, using therest 4 wells as control. Two glass slices were stuck with filter paper around to a thickness of 3mm and put into2 culture utensils, and inoculated with C6 and C6tk + cells respectively. Two days later, when 80% of cellswere confluent, the 2 glass slices were interchanged and added with the media containing 0. 5ug/ml GCV sepa-rately. The forms of cells were observed 6d later. Results C6 cell group showed bystander effect. However,Hela cell group showed no the effect. Most of the tk + cells in culture utensils were killed, while tk- cellsshowed no change. Conclusion Bystander effect is related to the gap junction during killing tumor cells byHSV - tk gene/GCV system.

关 键 词：HSV-TK基因 基因治疗 肿瘤 旁观者效应 缝隙连接 GCV系统 

分 类 号：R730.5[医药卫生—肿瘤] R394.8[医药卫生—临床医学]