顺铂对视网膜母细胞瘤细胞B7-H1表达的影响  

The influence of Cisplan on the expression of B7-H1 in retinoblastoma cells

作　　者：陈震[1] 邢怡桥[1] 徐冲[1] 龚文容[2] 

机构地区：[1]武汉大学人民医院眼科,430060 [2]武汉大学人民医院中心实验室,430060

出　　处：《中华眼底病杂志》2011年第1期25-28,共4页Chinese Journal of Ocular Fundus Diseases

摘　　要：目的 观察顺铂对视网膜母细胞瘤（RB）细胞B7-H1表达的影响,并探讨其作用机制.方法 分别采用浓度为0.375、0.750、1.500、3.000、6.000μg/ml顺铂处理人RB细胞HXO-Rb44细胞,作为不同浓度实验组;以RPMI 1640细胞培养液处理HXO-Rb14细胞作为空白对照组.采用逆转录-聚合酶链反应（RT-PCR）和荧光定量聚合酶链反应（PCR）检测HXO-Rb44细胞B7-H1 mRNA的表达;免疫荧光染色和流式细胞术检测HXO-Rb44细胞B7-H1蛋白表达.采用1.500μg/ml顺铂处理HXO-Rb44细胞0、15、30、60、120 min,蛋白免疫印迹（Western blot）检测细胞外信号调节激酶1/2（ERK1/2）的磷酸化水平.结果 0.375、0.750、1.500、3.000、6.000 μg/ml组HXO-Rb44细胞B7-H1 mRNA表达均较空白对照组高,差异有统计学意义（F=395.478,P=0.000）.0.375、0.750、1.500、3.000、6.000μg/ml组HXO-Rb44细胞B7-H1蛋白表达均较空白对照组高,差异有统计学意义（F=112.03,P=0.000）.Western blot检测显示,1.500μg/ml顺铂激活HXO-Rb44细胞ERK1/2蛋白的磷酸化水平;随时间延长,其磷酸化水平逐渐增高,至30 min达高峰,以后又逐步下降.结论 顺铂能促进RB细胞B7-H1 mRNA和蛋白的表达;ERK1/2信号通路可能参与了这一过程.Objective To observe the influence of cisplan on the expression of B7-H1 in retinoblastoma （RB） cells,and to investigate its mechanism. Methods Human RB cell line HXO-Rb44 cells were treated by 6 different concentrations of cisplan （0. 000, 0. 375, 0. 750, 1. 500, 3. 000, 6.000 μg/ml）,and their B7-H1 mRNA expression was determined by the reverse-transcription polymerase chain reaction （RT-PCR） and fluorescence quantitative PCR （FQ-PCR）;the B7-H1 protein expression was determined by immunofluorescence and flow cytometry. HXO-Rb44 cells were treated by 1.5 μg/ml cisplan for 0, 15, 30,60, 120 min, then the phosphorylation of extracellular signal-regulated kinase 1/2 （ERK1/2） was detected by Western blot. Results The expression of B7-H1 mRNA and protein in the 0. 375, 0. 750, 1. 500,3. 000, 6. 000 μg/ml group were significantly higher than that of the blank control group （F= 395. 478,112.03;P=0. 000）. Western blot showed that cisplan （1.5μg/ml） could activate ERK1/2 by increasing its phosphorylation in HXO-Rb44 cells. After cisplan treatment, the phosphorylation of ERK1/2 increased gradually and reached its peak at 30 min, and then went down gradually. Conclusion Cisplan can promote the expression of B7-H1 and activate ERK1/2 in RB cells.

关 键 词：视网膜母细胞瘤/药物疗法 顺铂/治疗应用 顺铂/投药和剂量 细胞外信号调节MAP激酶类 

分 类 号：R739.7[医药卫生—肿瘤]