CIB1下调表达对U87胶质瘤细胞增殖和细胞周期的影响  

Effects of CIB1 Down-expression on Cell Proliferation and Cell Cycle of U87 Glioma Cells

作　　者：惠玲[1] 闫蔚[1] 张富婷 王晓辉[1] 杨霄鹏[1] 马明仁[1] 贾庆华[1] 唐瑜[1] 马俊[1] 

机构地区：[1]兰州军区兰州总医院医学实验中心甘肃省干细胞与基因药物重点实验室,兰州730050

出　　处：《解放军医药杂志》2017年第10期10-15,共6页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科学基金(81372177);全军医药卫生科研基金目(CLZ12JA08);甘肃省自然科学基金(1107RJZA106,1308RJZA284)

摘　　要：目的探讨钙整合素结合蛋白1(CIB1)下调表达对U87胶质瘤细胞增殖和周期调节的作用。方法体外培养U87胶质瘤细胞,通过感染携带si CIB的p GV-si CIB1慢病毒获得CIB1下调表达的U87胶质瘤细胞,将细胞分为p GV-si CIB感染组、对照慢病毒感染组和对照组,采用甲基噻唑基四唑检测细胞增殖情况,流式细胞术检测各组细胞凋亡和细胞周期的改变,定量反转录聚合酶链反应、蛋白免疫印迹法检测U87胶质瘤细胞相关基因和蛋白表达情况。结果p GV-si CIB1感染U87细胞的最适感染复数值为5。si CIB1感染组CIB1 mRNA和蛋白表达低于对照慢病毒感染组和对照组(P<0.05)。p GV-si CIB1感染组FOXO1、MDM2 mRNA及Cyclin D1、Bcl-2 mRNA和蛋白、p-AKT蛋白表达水平低于对照慢病毒感染组,而Bax、p53、Caspase3 mRNA和蛋白表达水平高于对照慢病毒感染组(P<0.05,P<0.01)。p GV-si CIB1感染组U87胶质瘤细胞抑制率、凋亡率与对照慢病毒感染组相比显著增加,与对照组和对照慢病毒感染组相比,G0/G1期细胞增加,S期细胞减少(P<0.05)。结论 CIB1下调表达抑制胶质母细胞瘤的生长,促进细胞凋亡。AKT信号通路可能是CIB1发挥作用的途径之一,提示CIB1有可能作为胶质瘤靶向治疗的靶点。Objective To investigate effects of calcium-and integrin-binding protein （CIB1） down-regulation on cell proliferation and cell cycle of U87 glioma cells. Methods U87 glioma cells were cultured in vitro, and U87 glioma cells of CIB1 down-regulation were obtained by infecting pGV-siCIB1 lentivirus. The cells were divided into PGV-siCIB group, lentivirus infection group and control group. Conditions of cell proliferation were detected by methyl thiazolyl tet-razolium （ MTT） assay, and changes of cell apoptosis and cell cycle were detected by flow cytometry method, and mRNA expressions of related molecules were detected by quantitative reverse transcription polymerase chain reaction （ qRT-PCR） and protein western blotting. Results The best multiplicity of infection （ MOI） value of U87 cells by pGV-siCIB1 lenti-virus infection was 5. Compared with those in lentivirus infection and control groups, expressions of mRNA and protein of CIB1 were significantly decreased in pGV-siCIB1 （P〈0. 05）. In pGV-siCIB1 group, mRNA expressions of FOXO1 and MDM2, mRNA and protein expressions of CyclinD1, Bcl-2 and p-AKT protein were significantly decreased, while mRNA and protein expressions of Bax, p53 and Caspase3 were significantly increased in lentivirus infection group （ P〈0. 05, P〈0. 01）. The cell inhibition and apoptosis rates of U87 glioma cells in pGV-siCIB1 group were significantly higher than those in lentivirus infection group, and number of G0/G1 stage cells was significantly increased, while number of S stage cells was significantly decreased compared with those in lentivirus infection and control groups （P〈0. 05）. Conclusion

关 键 词：神经胶质瘤 钙整合素结合蛋白1 细胞增殖 细胞周期 

分 类 号：R730.264[医药卫生—肿瘤]