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目的:在TJ905胶质母细胞瘤细胞系中观察miR-146b-5p对MMP16的调控作用及其对细胞侵袭、迁移、增殖和凋亡的影响。方法:分别用无义序列表达质粒(对照组)和miR-146b-5p表达质粒(P-miR-146b-5p组)转染TJ905细胞,用qRT-PCR和Western blot检测两组细胞miR-146b-5p的表达水平及MMP16 mRNA和蛋白的表达水平,用体外迁移侵袭实验及流式细胞术检测转染细胞迁移、侵袭、细胞周期分布和凋亡水平,并分析这些变化的相互关系。结果:与对照组相比,p-miR-146b-5p组MMP16 mRNA和蛋白表达量明显降低,二者间呈正相关且均与miR-146b-5p表达量呈负相关。p-miR-146b-5p组侵袭和迁移细胞数均明显低于对照组,并均与同组MMP16蛋白表达量呈正相关;而凋亡水平明显高于对照组,并与同组miR-146b-5p表达量呈正相关,但两组细胞周期时相分布无显著性差异。结论:miR-146b-5p是胶质瘤的抑瘤miRNA;补充外源性miR-146b-5p可促进胶质瘤细胞凋亡,并通过抑制靶基因MMP16表达阻止其侵袭迁移;提示miR-146b-5p在恶性胶质瘤基因治疗方面具有重要的潜在应用价值。
OBJECTIVE: To observe the regulatory effect of miR-146b-5p on MMP16 and its effect on cell invasion, migration, proliferation and apoptosis in TJ905 glioblastoma cell line. Methods: TJ905 cells were transfected with nonsense sequence expression plasmid (control group) and miR-146b-5p expression plasmid (P-miR-146b-5p group). The expression of miR-146b was detected by qRT- -5p and the expression of MMP16 mRNA and protein. The migration, invasion, cell cycle distribution and apoptosis of transfected cells were detected by in vitro migration and invasion assay and flow cytometry, and the relationship between these changes was analyzed. Results: Compared with the control group, the expression of MMP16 mRNA and protein in p-miR-146b-5p group was significantly lower than that in the control group, which was negatively correlated with the expression of miR-146b-5p. The number of invasive and migrating cells in p-miR-146b-5p group was significantly lower than that in control group, and both were positively correlated with the expression of MMP16 protein in the same group, while the apoptosis level was significantly higher than that in control group, 5p expression was positively correlated, but the two groups of cell cycle phase distribution no significant difference. Conclusion: miR-146b-5p is a tumor suppressor miRNA of glioma. Supplementation of exogenous miR-146b-5p can promote glioma cell apoptosis and inhibit its invasion and migration by inhibiting the expression of target gene MMP16. It is suggested that miR-146b -5p has important potential value in gene therapy of malignant glioma.