三氧化二砷对人恶性黑素瘤A_(375)细胞抑制机制的探讨  

Research of the inhibitory mechanism of arsenic trioxide on human amalignant melanoma A_(375) cells

作　　者：黄桦[1] 高洁[2] 

机构地区：[1]蚌埠医学院生物化学与分子生物学教研室,安徽蚌埠233030 [2]蚌埠医学院第一附属医院检验科,安徽蚌埠233004

出　　处：《蚌埠医学院学报》2012年第11期1277-1279,1282,共4页Journal of Bengbu Medical College

基　　金：蚌埠医学院课题资助项目(BY0827)

摘　　要：目的:探讨三氧化二砷(As2O3)对人恶性黑素瘤A375细胞的生长抑制作用,以及对Caspase-3活性表达以及肿瘤细胞凋亡的影响,为临床治疗恶性黑素瘤提供理论和实验依据。方法:将不同浓度的As2O3作用于人恶性黑素瘤A375细胞,荧光染色观察A375细胞的凋亡变化,流式细胞术检测A375细胞的凋亡率;用免疫组织化学染色法检测不同浓度As2O3作用于A375细胞后Caspase-3蛋白的表达;半定量RT-PCR法检测Survivin蛋白的表达。结果:荧光染色后镜下可见As2O3可致A375细胞的细胞核呈致密浓染的不规则黄绿色颗粒状或块状荧光的凋亡现象;As2O3能诱导A375细胞的凋亡,用5、10、20μmol/L的As2O3诱导A375细胞24 h:检测其细胞的凋亡率依次为7.07%、38.66%和4.34%,可诱导Caspase-3蛋白的表达,抑制Survivin蛋白的表达。结论:As2O3能致A375细胞形态改变,诱导细胞凋亡;As2O3可诱导A375细胞表达Caspase-3蛋白,抑制Survivin蛋白的表达。Objective：To investigate the growth inhibition role of arsenic trioxide（As2O3） on human amalignant melanoma A375 cell,and observe whether Caspase-3 protein can be expressed in A375 cell after induced by As2O3.To provide theory and experiment base for treament of malignant melanoma. Methods：The morphologic changes of A375 cells were detected by fluorescence staining,apoptosis rate of A375 cells was detected with flow cytometry.Caspase-3 protein was detected by immunehistochemistry in A375 cells.The expression of Survivin mRNA was examined by semi-quantitative RT-PCR. Results：As2O3 caused morphologic change of A375 cells by fluorescence staining.As2O3 promoted A375 cell apoptosis significantly.When A375 cells were treated with As2O3 in 5,10,and 20 μmol/L for 24 hours,the apoptosis rate was 7.07%,38.66% and 4.34%.Caspase-3 protein was expressed positively in A375 cells by co-culture with As2O3.The expression of Survivin mRNA was inhibited by As2O3. Conclusions：As2O3 can cause morphologic change of A375 cells and promote apoptosis happening.As2O3 can induce A375 cells to express Caspase-3 protein.As2O3 can inhibit Survivin protein expression in A375 cells.

关 键 词：黑素瘤 三氧化二砷 细胞凋亡 CASPASE-3 SURVIVIN A375细胞 

分 类 号：R739.5[医药卫生—肿瘤]