摘要
目的：探讨抑癌基因 RUNX3对胶质瘤细胞迁移和侵袭的影响及其机制。方法应用脂质体2000将真核表达质粒pFlag－RUNX3和pFlag－Control分别转入U251和U87细胞中，Western blot技术检测RNUX3蛋白表达，通过迁移实验、人工基底膜细胞侵袭实验来检测细胞的迁移和侵袭能力，Western blot和明胶酶谱实验检测MMP－2的蛋白表达和酶活性。结果与对照组相比，转染pFlag－RUNX3质粒24 h后U251和U87细胞中RUNX3蛋白表达显著增高；细胞迁移能力分别下降了86％和74％；细胞侵袭能力下降了83％和70％，差异均有统计学意义（P＜0．01）；转染RUNX3可以明显抑制2种胶质瘤细胞MMP－2蛋白的表达和酶活性。结论RUNX3可以通过MMP－2途径抑制胶质瘤细胞迁移和侵袭能力。
Objective To investigate the effects of RUNX3 on the migration and invasion of human glioma cells and possible mechanism.Methods The plasmids pFlag-RUNX3 and pFlag-Control were transfected into U251 and U87 glioma cells using Lipofectamine 2000 reagent, respectively.The expression of RUNX3 in both glioma cells were meas-ured by Western blotting.The effects of RUNX3 on the migration and invasion of these glioma cells were assessed by mi-gration assay and matrigel cell invasion assay.Also, the activities and expression of MMP-2 were measured by gelatin zymography and Western blot, respectively.Results Compared to the control, transfection of pFlag-RUNX3 plasmids could markedly enhance the amounts of RUNX3 in U251 and U87 cells.In cell migration assay, RUNX3 inhibited the in-vasive abilities of U251 and U87 cells by 86%and 74%, respectively （P〈0.01）.RUNX3 inhibited the invasive abili-ties of U251 and U87 cells in matrigel-coated Boyden chamber by 83%and 70%, respectively （P〈0.01）.Transfec-tion of pFlag-RUNX3 also suppressed the activities and expression of MMP-2.Conclusion RUNX3 may weaken the migration and invasion of glioma cells through inhibiting MMP-2 pathway.
出处
《徐州医学院学报》
CAS
2015年第10期644-647,共4页
Acta Academiae Medicinae Xuzhou
基金
徐州市科技项目（XM138075）
