【摘要】 目的探讨变异型IκBα(IκBαM)基因对人类多形性胶质母细胞瘤(GBM)细胞中基质金属蛋白酶(MMP)-2、-9表达的调控作用。方法通过构建质粒、基因转染以及IκBαM蛋白表达的筛选,建立稳定表达IκBαM的人类GBM细胞株,运用逆转录-聚合酶链反应(RT-PCR)技术检测细胞及肿瘤组织内MMP-2、MMP-9在RNA水平的表达;并将肿瘤细胞植入裸鼠皮下制作异位移植瘤生长动物模型,进一步通过免疫组织化学染色方法分析MMP-2、MMP-9的蛋白表达。结果RT-PCR结果显示,体外试验中,MMP-2与对应GAPDH平均吸光度值的比值分别为1.450±0.180 (G36△-W)、0.292±0.040(G36△-M)、1.187±0.140(G36△-P)和1.463±0.160(G36△);MMP-9与对应GAPDH平均吸光度值的比值分别为1.424±0.130(G36△-W)、0.275±0.020(G36△-M)、1.357±0.180(G36△-P)和1.608±0.240(G36△)。在体内试验中,MMP-2与对应GAPDH平均吸光度值的比值分别为0.870±0.060(G36△-W)、0.024±0.010(G36△-M)、0.785±0.070(G36△-P)和0.686±0.070(G36△);MMP-9与对应GAPDH平均吸光度值的比值分别为0.768±0.010(G36△- W)、0.054±0.010(G36△-M)、0.802±0.020(G36△-P)和0.746±0.020(G36△)。在体外和体内试验中,G36△-M组与其余各组之间的差异均有统计学意义(P<0.05),而其他3组间差异无统计学意义。体内试验肿瘤组织中,MMP-2及MMP-9的在G36△-M组中的染色显著低于G36△、G36△-W和G36△-P组;而在G36△、G36△-W和G36△-P三组间的表达差异无统计学意义。结论IκBαM基因可以在分子及蛋白水平显著抑制人类恶性胶质瘤中MMP-2、MMP-9的表达,减弱肿瘤细胞的侵袭和浸润能力。更多还原

【Abstract】 Objective To investigate the regulation of mutant-type IκBα(IκBαM)to the expres- sion of matrix metalloproteinases(MMP-2 and MMP-9)in the human glioblastoma multiform(GBM) cells.Methods Human GBM cell line was used,transfected with IκBαM gene and the selection of IκBαM protein’s expression,to establish cell line which steadily express IκBαM protein.Then total RNA were isolated and analyzed the expression of MMP-2 and MMP-9 by RT-PCR method at RNA level;and the cells were injected subcutaneously into the nude mice,then compare the expression of MMP-2 and MMP-9 by immunohistochemistry method.Results The RT-PCR results show that in vitro MMP-2/GAPDH(ratio of average optic density)are respectively 1.450±0.180(G36△-W),0.292±0.040(G36△-M),1.187±0.140(G36△-P)and 1.463±0.160(G36△);and MMP-9/GAPDH are respectively 1.424±0.130 (G36△-W),0.275±0.020(G36△-M),1.357±0.180(G36△-P)and 1.608+0.240(G36△);in vivo MMP-2/GAPDH are respectively 0.870±0.060(G36△-W),0.024±0.010(G36△-M),0.785±0.070 (G36△-P)and 0.686±0.070(G36△);and MMP-9/GAPDH are respectively 0.768±0.010(G36△- W),0.054±0.010(G36△-M),0.802±0.020(G36△-P)and 0.746±0.020(G36△)in vivo.The differences between G36△-M group and each other group all have statistical significance(P<0.05)in vitro and in vivo,but the differences among the other three groups(G36△-W,G36△-P and G36△)have no statistical significance.The stain of MMP-2 or MMP-9 protein was decrease significantly in G36△-M group tumor tissues in vivo,but the other three groups(G36△-W,G36△-P and G36△)almost same.Conclusion IκBαM gene could inhibit the expressions of MMP-2 and MMP-9 in the human GBM cell line or in the tumor tissues both in mRNA and protein level,reduce the invasion and infiltrate ability of tumor cells.更多还原