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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 the effect of CIB1 down-regulation on the proliferation and cycle regulation of U87 glioma cells. Methods U87 glioma cells were cultured in vitro and U87 glioma cells down-regulated by CIB1 were obtained by infecting pGV-si CIB1 lentivirus carrying si CIB. The cells were divided into pGV-si CIB infected group and control lentiviral infected group And control group, the cell proliferation was detected by methylthiazolyl tetrazolium, the apoptosis and cell cycle were detected by flow cytometry, quantitative reverse transcriptase polymerase chain reaction and Western blotting were used to detect the expression of U87 glioma Cell-related genes and protein expression. Results The optimal infection multiplicity of infection of U87 cells by pVV-si CIB1 was 5. The expression of CIB1 mRNA and protein in si CIB1 infection group was lower than that in control lentivirus infection group and control group (P <0.05). The expression of FOXO1, MDM2 mRNA, Cyclin D1, Bcl-2 mRNA and protein, p-AKT protein in GV-si CIB1 infection group was lower than that in control lentivirus group, while the expression of Bax, p53, Caspase3 mRNA and protein were higher Lentivirus infection group (P <0.05, P <0.01). Compared with the control group and the control lentivirus group, the inhibitory rate and apoptosis rate of U87 glioma cells in pGV-si CIB1 infection group were significantly increased compared with the control group and control lentiviral infection group, cells in G0 / G1 phase increased, S phase Cells decreased (P <0.05). Conclusion The down-regulation of CIB1 inhibits the growth of glioblastoma cells and promotes apoptosis. AKT signaling pathway may be one of the ways CIB1 play a role, suggesting that CIB1 may serve as a target for targeted treatment of glioma.