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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 4
Objective To investigate the bystander effect of herpes simplex virus thymidine kinase / ganciclovir (HSV TK / GCV) in the treatment of ovarian cancer and its relationship with the expression of connexin (Cx) 4 3, and to observe the effect of all-trans retinoic acid (RA) on the relationship between the two. Methods MTT method was used to compare the bystander effect of HSV TK / GCV treatment with OVCAR3 and CAOV3 cells before and after treatment with RA. The effects of HSV TK / GCV on the bystander effect were compared by Western Blot ), Indirect immunofluorescence staining before and after the two ovarian cancer cell lines RA Cx4 3 expression, Cx4 3 expression in mean fluorescence intensity. Results (1) HSV TK / GCV had obvious bystander effect on OVCAR3 cells and weak onstander effect on CAOV3 cells, the difference was significant (P <0.05). (2) RA enhanced the bystander effect of OVCAR3 cells (P <0 05), but did not affect the bystander effect of CAOV3 cells (P 0 05). (3) The results of flow cytometry showed that the expression of Cx4 3 in OVCAR3 cells was 445, while the expression of Cx4 3 in CAOV3 cells was 0 89. The difference was significant (P <0 05). Western Blot, indirect immunofluorescence staining results also showed that OVCAR3 cells have Cx4 3 expression, and located in the cell membrane, and CAOV3 cells without Cx4 3 expression. (4) After RA treatment, Western Blot and indirect immunofluorescence staining showed that the expression of Cx4 3 in both ovarian cancer cell lines was increased. The results of flow cytometry showed that the expression of Cx4 3 was significantly increased in OVCAR3 cells,