干扰BAG-1表达对黑素瘤B16F10细胞放射敏感性的影响  

Effect of inhibiting of BAG-1 gene expression on radiosensitivity of melanoma B16F10 cells

作　　者：金德蕙[1] 王占想[2] 宋亚丽[1] JIN Dehui;WANG Zhanxiang;SONG Yali(Shandong Provincial Hospital,Jinan 250021,China;Henan Puyang Oilfield General Hospital,Puyang 457000,China)

机构地区：[1]山东省立医院皮肤科,济南250021 [2]河南濮阳市油田总医院皮肤科,濮阳457000

出　　处：《中国麻风皮肤病杂志》2018年第12期708-711,共4页China Journal of Leprosy and Skin Diseases

基　　金：山东省自然科学基金青年基金(编号:ZR2011HQ056);山东省重点研发计划(编号:2015GSF118128)

摘　　要：目的:明确BAG-1基因表达对黑素瘤B16F10细胞放射敏感性的影响及机制。方法:B16F10细胞株分为未转染组(Control组)、阴性对照质粒转染组(NC-shRNA组)和转染BAG-1-shRNA组(BAG-1-shRNA组)。克隆计数法计算克隆形成率和存活分数,描绘细胞存活曲线。流式细胞仪检测细胞凋亡率,Western blot检测BAG-1沉默对黑素瘤细胞凋亡相关蛋白Caspase3、8、9、PARP表达。结果:X线8Gy剂量照射后,Control组、NC-shRNA组和BAG-1-shRNA组细胞的存活率分别为1.9%,1.75%和0.32%,细胞凋亡率分别为7. 1%,9. 2%和25. 3%,差异具有统计学意义(均P <0. 05)。BAG-1-shRNA组凋亡蛋白c-PARP及c-Caspase 3、Caspase 8、Caspase 9水平分别为0.36,0.37,0.35和0.30,高于control组(0.10,0.04,0.11,0.2)和NC-shRNA组(0.13,0.12,0.20,0.26)。结论:干扰BAG-1基因表达可显著增高黑素瘤B16F10细胞对X放射线的敏感性,促进细胞凋亡可能是放射增敏的机制之一。Objective:To determine effect of inhibiting of BAG-1 gene expression on radiosensitivity of melanoma B16F10 cells and explore the mechanism.Methods:B16F10 cells were divided into three groups:control group,NC-shRNA group and BAG-1-shRNA group.Medical linear accelerator irradiation and clone counting method were used to calculate the clone formation rate and survival fraction,and the cell survival curve was delineated.The apoptosis rates were detected by flow cytometry.The expression level of apoptosis-related proteins Caspase 3,8,9,and c-PARP in melanoma cells were detected by Western blot.Results:After X-ray 8Gy dose irradiation,the survival rates of cells in the control group,NC-shRNA group and BAG-1-shRNA group linear were 1.9%,1.75%,0.32%and the apoptosis rate were 7.1%,9.2%and 25.3%respectively,with significant differences(P<0.05).The levels of c-PARP,Caspase 3,8,9 were 0.36,0.37,0.35 and 0.30 respectively,which were higher than those in the control group and NC-shRNA group,with significant differences(P<0.05).Conclusion:RNA interference of BAG-1 gene expression can significantly increase the sensitivity of melanoma cells to X-ray radiation,which may be associated with accelerating cells apoptosis.

关 键 词：黑素瘤 BAG-1 RNA干扰 放射增敏 细胞凋亡 

分 类 号：R739.5[医药卫生—肿瘤]