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目的观察微小RNA-15b(miRNA-15b)对人脑胶质瘤细胞株U87细胞侵袭能力的影响。方法体外培养U87细胞,分为空白对照组、阴性对照组和miRNA-15b寡聚核苷酸拟似物处理组(miRNA-15b组)。实时荧光定量PCR检测miRNA-15b表达水平,Transwell小室实验检测细胞侵袭能力,Western blot实验检测细胞侵袭相关蛋白Ras相关C3内毒素底物1(Rac1)和Rac3表达水平。结果脂质体转染U87细胞48h后,miRNA-15b组细胞miRNA-15b相对表达量高于阴性对照组和空白对照组(18.47±2.31vs.12.75±1.98和11.22±1.32)(P<0.05);miRNA-15b组细胞穿过人工基底膜数少于阴性对照组和空白对照组[(11.0±1.8)个vs.(34.0±5.1)个和(32.0±4.8)个](P<0.05);miRNA-15b组U87细胞中Rac1和Rac3蛋白表达水平较阴性对照组和空白对照组下降(P<0.05)。结论 miRNA-15b可通过下调Rac1和Rac3蛋白表达抑制U87细胞的侵袭能力。
Objective To investigate the effect of microRNA-15b (miRNA-15b) on invasion of human glioma U87 cells. Methods U87 cells were cultured in vitro and divided into blank control group, negative control group and miRNA-15b oligonucleotide mimic group (miRNA-15b group). Real-time fluorescence quantitative PCR was used to detect the expression of miRNA-15b. Transwell assay was used to detect the invasion ability of cells. Western blot was used to detect the expression of ras3, Rac1 and Rac3. Results Compared with the negative control group and the blank control group, the miRNA-15b expression level in the miRNA-15b group was significantly higher than that in the untreated U87 cells (18.47 ± 2.31 vs. 12.75 ± 1.98 and 11.22 ± 1.32, P <0.05) (11.0 ± 1.8) vs (34.0 ± 5.1) and (32.0 ± 4.8), respectively (P <0.05) in the miRNA-15b group compared with the negative control group and the blank control group; The expression of Rac1 and Rac3 in the miRNA-15b group was lower than that in the negative control group and the blank control group (P <0.05). Conclusion miRNA-15b can inhibit the invasion of U87 cells by downregulating the expression of Rac1 and Rac3.