摘要
目的 探讨单纯疱疹病毒 胸苷激酶 /更昔洛韦 (HSV TK/GCV)在卵巢癌治疗中的旁观者效应及其与间隙连接蛋白 (Cx) 4 3表达的关系 ,同时观察全反式维甲酸 (RA)对两者关系的影响。方法 四甲基偶氮唑蓝 (MTT)法比较经HSV TK/GCV治疗及RA作用前后卵巢癌细胞株OVCAR3、CAOV3细胞旁观者效应的强弱 ;采用流式细胞仪 ,蛋白质免疫印迹法 (WesternBlot)、间接免疫荧光染色检测两卵巢癌细胞株RA作用前后Cx4 3的表达 ,Cx4 3的表达以平均荧光强度表示。结果 ( 1)HSV TK/GCV对OVCAR3细胞产生较明显的旁观者效应 ,而对CAOV3细胞旁观者效应弱 ,两者比较 ,差异有显著性 (P <0 0 5 )。( 2 )RA对OVCAR3细胞的旁观者效应有增强作用 (P <0 0 5 ) ,但不影响CAOV3细胞的旁观者效应 (P >0 0 5 )。( 3 )流式细胞仪检测提示 ,OVCAR3细胞中Cx4 3的表达为 4 4 5 ,而CAOV3细胞中Cx4 3的表达为 0 89,两者比较 ,差异有显著性 (P <0 0 5 ) ;WesternBlot、间接免疫荧光染色检测结果也表明 ,OVCAR3细胞有Cx4 3的表达 ,且定位于细胞膜 ,而CAOV3细胞中无Cx4 3的表达。 ( 4 )RA作用后 ,WesternBlot、间接免疫荧光染色检测均显示 ,两卵巢癌细胞株的Cx4 3表达增加 ;流式细胞仪检测提示 ,细胞中Cx4 3表达均明显增加 ,OVCAR3细胞由 4
Objective To explore the relationship of connexin 43(Cx43) and bystander effect in ovarian tumor cells in herpes simplex virus thymidine kinase/ganciclovir (HSV TK/GCV) gene therapy in vitro, and to investigate the effection of all trans retinoic acid (RA) on expression of Cx43 and bystander effect Methods Cx43 expression was examined with flowcyto metry, Western Blot, and immunofluorescence in two ovarian tumor cells OVCAR3, CAOV3 before and after RA treatment Bystander effect was determined by the cells growth inhibitory rate with methyl thiazolyl tetrazolium Results Following exposure to ganciclovir, there was much greater bystander killing in OVCAR3 than in CAOV3 ( P <0 05) The expression of Cx43 was detected in OVCAR3 with flowcytometry and Wstern Blot, but it could not be detected in CAOV3 The expression of Cx43 in both cell lines could be induced by RA Immunofluoresence staining showed that OVCAR3 Cx43 protein is located in membrane surface, whereas CAOV3 is in cytoplasm RA could not change the location of Cx43 protein in both cell lines Conclusions There is relationship between Cx43 expression and HSV TK/GCV bystander effect. HSV TK/GCV bystander effect can be inhanced by RA in ovarian cancer
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2001年第9期542-545,共4页
Chinese Journal of Obstetrics and Gynecology
基金
国家自然科学基金资助项目 ( 39840 0 0 9
39870 765 )
国家杰出青年科学基金资助项目 ( 30 0 2 5 0 17)
