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目的:研究microRNA-10a(miR-10a)对人脑胶质瘤细胞系U87MG侵袭性的影响。方法:脂质体包被miR-10a反义寡聚核苷酸(miR-10a antisense oligodeoxynucleotide,miR-10a-anti-ODN),转染胶质瘤U87MG细胞,并设无义miRNA转染组和空白对照组,流式细胞术和荧光显微镜检测miR-10a-anti-ODN对U87MG细胞的转染效率,流式细胞术检测转染miR-10a-anti-ODN后U87MG细胞的凋亡和细胞周期,MTT法检测U87MG细胞的增殖,Transwell实验检测miR-10a-anti-ODN对U87MG细胞迁移和侵袭的影响;RT-PCR和Western blotting法分别检测U87MG细胞中MMP-2、MMP-9、MMP-14 mRNA及蛋白的表达。结果:转染miR-10a-anti-ODN后,U87MG细胞的增殖、周期和凋亡无明显改变,U87MG细胞的侵袭[(87±7.1)vs(155±3.7)、(149±6.6)个细胞,P<0.05]和迁移[(78.0±5.2)vs(150.3±3.7)、(147.3±6.6)个细胞,P<0.05]能力明显下降,侵袭相关因子MMP-2、MMP-9、MMP-14的mRNA及蛋白表达水平也明显下降。结论:miR-10a通过调控MMP-2、MMP-9、MMP-14的表达促进胶质瘤U87MG细胞的侵袭,miR-10a可能是胶质瘤治疗的潜在靶点。
AIM: To investigate the effect of microRNA-10a (miR-10a) on the invasiveness of human glioma cell line U87MG. METHODS: The miR-10a antisense oligodeoxynucleotide (miR-10a antisense oligodeoxynucleotide) was transfected into glioma U87MG cells by Lipofectamine 2000 and transfected with nonsense miRNA The transfection efficiency of miR-10a-anti-ODN on U87MG cells was detected by flow cytometry and fluorescence microscopy. The apoptosis and cell cycle of U87MG cells transfected with miR-10a-anti-ODN were detected by flow cytometry The effect of miR-10a-anti-ODN on the migration and invasion of U87MG cells was detected by Transwell assay. The expressions of MMP-2, MMP-9 and MMP-9 in U87MG cells were detected by RT- 14 mRNA and protein expression. Results: The proliferation, cell cycle and apoptosis of U87MG cells were not significantly changed after transfection with miR-10a-anti-ODN. The invasion of U87MG cells [(87 ± 7.1) vs (155 ± 3.7), (149 ± 6.6) , P <0.05] and migration [(78.0 ± 5.2) vs (150.3 ± 3.7) and (147.3 ± 6.6) cells, respectively, P <0.05]. The invasion-related factors MMP-2, MMP-9 and MMP- MRNA and protein expression levels also decreased significantly. Conclusion: miR-10a can promote the invasion of glioma U87MG cells through regulating the expression of MMP-2, MMP-9 and MMP-14. MiR-10a may be a potential therapeutic target for glioma.