腺病毒介导PTEN和TIMP-2抑制胶质瘤U87体外侵袭力的研究

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

SUPPRESSION EFFECT OF ADENOVIRUS-MEDIATED GENE TRANSFER OF PTEN AND TIMP-2 ON INVASION OF HUMAN U87 GLIOMA CELLS IN VITRO

Objective To evaluate the effects of adenovirus mediated gene transfer of TIMP 2 and PTEN on invasion of human U87 glioma cells in vitro . Methods U87 cells were transinfected with AdTIMP 2 and AdPTEN in vitro . The mRNA and protein expressions of TIMP 2 and PTEN were detected with RT PCR and Western blotting, respectively. The relative activity of MMP 2 and MMP 9 was determined by gelatin zymogram, and invasion of U87 in vitro was observed using Boyden chamber. Results Gene and protein expressions of PTEN and TIMP 2 were shown to be up regulated when U87 was transinfected with AdPTEN and AdTIMP 2. The number of invasion cells of U87 infected with AdX gal, AdPTEN, AdTIMP 2 and PTEN TIMP 2 was 55 64 13 27, 48 26 14 75, 35 27 10 94, 27 38 12 81, and 19 16 5 45, respectively. In vitro invasion of glioma cells was significantly inhibited after infected with AdTIMP 2 and/or AdPTEN, while the inhibition effect was more remarkable in the combined group than that in single group, and it was not consistent with the change in MMPs activity. Conclusion These results imply that combined TIMP 2 and PTEN gene therapy mediated by adenorirus may be useful for anti invasion therapy of malignant glioma