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目的 探讨腺病毒介导PTEN和TIMP 2对人脑胶质瘤细胞体外侵袭力的抑制作用。方法 用含PTEN和TIMP 2基因的重组腺病毒载体体外转染人胶质瘤细胞系U87,用RT PCR、Westernblot检测目的基因与蛋白表达。通过Boyden小室法检测U87转染前后细胞体外侵袭力的变化 ,同时采用胶原酶谱法分析MMP 2、MMP 9酶的相对活性。结果 转染AdPTEN、AdTIMP 2后U87细胞中目的基因与蛋白的表达上调 ,U87、AdX gal、AdPTEN、AdTIMP 2和PTEN +TIMP 2的侵袭细胞数分别为 5 5 6 4± 13 2 7、4 8 2 6± 14 75、35 2 7± 10 94、2 7 38± 12 81和 19 16± 5 4 5 ,转染靶基因后U87细胞的体外侵袭力受到明显抑制 ,联合转染后侵袭力的抑制更明显 ,这与MMP的改变并不完全一致。结论 重组腺病毒载体介导的PTEN与TIMP 2联合基因治疗可能是抗胶质瘤侵袭的一个有用方法
Objective To investigate the inhibitory effect of adenovirus-mediated PTEN and TIMP 2 on invasiveness of human glioma cells in vitro. Methods The human glioma cell line U87 was transfected with the recombinant adenovirus vector containing PTEN and TIMP 2 gene in vitro. The expression of target gene and protein was detected by RT PCR and Western blot. The in vitro invasiveness of U87 transfected U87 cells was detected by Boyden chamber method, and the relative activities of MMP2 and MMP9 were analyzed by collagenase method. Results The expression of target gene and protein in U87 cells was up-regulated after AdPTEN and AdTIMP 2 transfection. The number of invasive cells of U87, AdX gal, AdPTEN, AdTIMP 2 and PTEN + TIMP 2 were respectively 5 5 6 4 ± 13 2 7 48 2 6 ± 14 75,35 2 7 ± 10 94,2 7 38 ± 12 81 and 19 16 ± 5 4 5, the invasion of U87 cells was significantly inhibited after transfected with the target gene, and the invasiveness of the transfected cells was inhibited More obviously, this is not exactly the same as the change of MMP. Conclusions Recombinant adenovirus vector mediated PTEN and TIMP 2 gene therapy may be a useful method for anti-glioma invasion