8-Br-cAMP提高HSV-TK/GCV自杀基因系统旁观者效应的研究

目的 探讨8-Br-cAMP对单纯疱疹病毒胸苷激酶（HSV-TK）/丙氧鸟苷（GCV）系统旁观者效应的增强作用及机理。方法 应用FCM、透射电镜检测8-Br-cAMP对口腔鳞癌细胞（Tca8113）、间隙连接蛋白43（Cx43）及间隙连接的诱导分化作用;采用析因分析法分析8-Br-cAMP对HSV-TK/GCV系统旁观者效应的影响。结果 口腔鳞癌细胞经 10-4mol/L 8-Br-CAMP处理后:FCM检测显示 Cx43表达阳性的细胞由治疗前的（4.8 ± 0.26）％上升到（21.3±0.65）％,两组比较P<0.001;透射电镜显示细胞超微结构出现分化,细胞间出现间隙连接;MTT检测显示增强了HSV-TK/GCV系统治疗时的旁观者效应,两者间存在协同关系。结论8-Br-cAMP可诱导分化口腔鳞癌细胞并显著提高HSV-TK/GCV系统治疗时的旁观者效应。

8-Br-cAMP AUGMENT THE BYSTANDER EFFECT OF HERPES SIMPLEX VIRUS THYMIDINE KINASE/GANCICLOVIR SYSTEM IN THE TREATMENT OF ORAL SQUAMOUS CARCINOMA CELL LINE

ve To investigate the augmentation effect and mechanism of 8-Br-cAMP (8-Bromo3:5-cyclic adeno-sine monophosphate) on the bystander effect of herpes simplex virus thymidine kinase /ganciclovir system in the treatment of oral squamous carcinoma cell line (Tea8113). Methods Flow cytometry (FCM) and transmission electron microscope were used to analyze the expression of Cx43 protein and formation of gap junctions in Tca8113 cells after treated with 8-Br-cAMP; The bystander effects of HSV-TK/GCV system before and after treated with 8-Br-cAMP were detected by MTT assays, the interaction was analyzed by factorial experiment. Results After treated with 8-Br-cAMP (10-4mol/L): the positive rate of Cx43 protein increased from 4.8 ± 0.26% to 21.3 ± 0.65% (P < 0.001); A series of cell morphological changes was observed , such as more tonofilaments and well-differentiated organelle; and gap junction was formed. After combined with 8-Br-cAMP, the bystander effect was significant improved (P <0.05). There had cooperation effect between 8-Br-cAMP and bystander effect. Conclusion 8-Br-cAMP can augment the bystander effect of HSV-TK/GCV system in the treatment of oral squamous carcinoma cell line. The mechanism might be due to up-regul ation of Cx43 gene and restore gap junction.