miR-128对胶质母细胞瘤U87细胞侵袭性的影响及作用机制

摘要

目的 探讨miR-128对胶质母细胞瘤U87细胞侵袭性的影响及其作用机制.方法 采用RT-PCR法测定胶质母细胞瘤U87细胞和SHG-44细胞中miR-128的表达情况,应用Transwell侵袭和迁移实验检测U87细胞的侵袭性,采用荧光素酶报告基因分析法检测miR-128调节的靶基因,应用Western blot分析法检测MMP-7蛋白的表达情况,并与阴性对照组(NC组)进行比较.结果 miR-128在SHG-44细胞、U87细胞中的表达量分别为100±0.13、0.52±0.16,差异有统计学意义(P<0.05).与NC组比较,肿瘤细胞高表达miR-128时两组穿膜细胞数分别为445±23、313±59,差异有统计学意义(P<0.05).与NC组比较,肿瘤细胞过度表达miR-128时,MMP-7蛋白表达明显减少,差异有统计学意义(P<0.05).结论 miR-128通过负性调控MMP-7蛋白的表达以抑制U87细胞的侵袭,提示miR-128可作为临床靶向治疗胶质母细胞瘤的关键.%Objective To investigate effects of miR-128 on invasion of glioblastoma U87 cells and its molec-ular mechanism. Methods Reverse transcription-quantitative polymerase chain reaction ( RT-PCR ) was used to measure the expression levels of miR-128 in glioblastoma U87 cells and SHG-44 cells. Migration and invasion of U87 cells were analyzed by transwell migration and invasion assays. Luciferase reporter assay was employed to detect cell target genes of miR-128 , while western blot analysis was performed to verify the expression of MMP-7 . Results The results showed that the expressions of miR-128 in SHG-44 cells and U87 cells were 1.00 ± 0.13 and 0.52 ± 0.16 re-spectively, and the difference was statistically significant (P<0.05). Compared with the NC group, the number of invasive cells was 445 ±23 and 313 ±59 respectively, and the difference was statistically significant (P<0.05). Compared with the NC group, when the tumor cells overexpressed miR-128, the expression of MMP-7 protein de-creased significantly, revealing significant difference (P<0.05). Conclusion The miR-128 could inhibit tumor invasion and migration in U87 cells by negatively regulating MMP-7 protein and may be identified as a potential thera-peutic target of glioblastoma.