中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2011年
1期
25-28
,共4页
陈震%邢怡桥%徐冲%龚文容
陳震%邢怡橋%徐遲%龔文容
진진%형이교%서충%공문용
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
視網膜母細胞瘤/藥物療法%順鉑/治療應用%順鉑/投藥和劑量%細胞外信號調節MAP激酶類
시망막모세포류/약물요법%순박/치료응용%순박/투약화제량%세포외신호조절MAP격매류
Retinoblastoma/drug therapy%Cisplatin/therapeutic use%Cisplatin/administration & dosage%Extracellular signal-regulated MAP kinases
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.
目的 觀察順鉑對視網膜母細胞瘤(RB)細胞B7-H1錶達的影響,併探討其作用機製.方法 分彆採用濃度為0.375、0.750、1.500、3.000、6.000μg/ml順鉑處理人RB細胞HXO-Rb44細胞,作為不同濃度實驗組;以RPMI 1640細胞培養液處理HXO-Rb14細胞作為空白對照組.採用逆轉錄-聚閤酶鏈反應(RT-PCR)和熒光定量聚閤酶鏈反應(PCR)檢測HXO-Rb44細胞B7-H1 mRNA的錶達;免疫熒光染色和流式細胞術檢測HXO-Rb44細胞B7-H1蛋白錶達.採用1.500μg/ml順鉑處理HXO-Rb44細胞0、15、30、60、120 min,蛋白免疫印跡(Western blot)檢測細胞外信號調節激酶1/2(ERK1/2)的燐痠化水平.結果 0.375、0.750、1.500、3.000、6.000 μg/ml組HXO-Rb44細胞B7-H1 mRNA錶達均較空白對照組高,差異有統計學意義(F=395.478,P=0.000).0.375、0.750、1.500、3.000、6.000μg/ml組HXO-Rb44細胞B7-H1蛋白錶達均較空白對照組高,差異有統計學意義(F=112.03,P=0.000).Western blot檢測顯示,1.500μg/ml順鉑激活HXO-Rb44細胞ERK1/2蛋白的燐痠化水平;隨時間延長,其燐痠化水平逐漸增高,至30 min達高峰,以後又逐步下降.結論 順鉑能促進RB細胞B7-H1 mRNA和蛋白的錶達;ERK1/2信號通路可能參與瞭這一過程.
목적 관찰순박대시망막모세포류(RB)세포B7-H1표체적영향,병탐토기작용궤제.방법 분별채용농도위0.375、0.750、1.500、3.000、6.000μg/ml순박처리인RB세포HXO-Rb44세포,작위불동농도실험조;이RPMI 1640세포배양액처리HXO-Rb14세포작위공백대조조.채용역전록-취합매련반응(RT-PCR)화형광정량취합매련반응(PCR)검측HXO-Rb44세포B7-H1 mRNA적표체;면역형광염색화류식세포술검측HXO-Rb44세포B7-H1단백표체.채용1.500μg/ml순박처리HXO-Rb44세포0、15、30、60、120 min,단백면역인적(Western blot)검측세포외신호조절격매1/2(ERK1/2)적린산화수평.결과 0.375、0.750、1.500、3.000、6.000 μg/ml조HXO-Rb44세포B7-H1 mRNA표체균교공백대조조고,차이유통계학의의(F=395.478,P=0.000).0.375、0.750、1.500、3.000、6.000μg/ml조HXO-Rb44세포B7-H1단백표체균교공백대조조고,차이유통계학의의(F=112.03,P=0.000).Western blot검측현시,1.500μg/ml순박격활HXO-Rb44세포ERK1/2단백적린산화수평;수시간연장,기린산화수평축점증고,지30 min체고봉,이후우축보하강.결론 순박능촉진RB세포B7-H1 mRNA화단백적표체;ERK1/2신호통로가능삼여료저일과정.
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.
