miR-148b-3p通过调控Wnt信号通路对胶质瘤细胞侵袭和迁移的影响  

Effect of miR-148b-3p on invasion and migration of glioma cells by regulating Wnt signaling pathway

作　　者：许辉[1] 沈杰 谢兰军[3] Xu Hui;Shen Jie;Xie Lanjun(Department of Neurology,Fuyang People's Hospital,Fuyang 236000,China;Department of Neurosurgery Xuanwu Hospital Affiliated to Beijing Capital Medical University,Beijing 100053,China;Department of Neurosurgery,Fuyang People's Hospital,Fuyang 236000,China)

机构地区：[1]阜阳市人民医院神经内科,236000 [2]首都医科大学附属宣武医院神经外科,北京100053 [3]阜阳市人民医院神经外科,236000

出　　处：《中华肿瘤杂志》2020年第7期565-569,共5页Chinese Journal of Oncology

基　　金：阜阳市卫生健康委科研立项(FY2019-006)。

摘　　要：目的:探讨miR-148b-3p对胶质瘤细胞侵袭和迁移的影响及其作用机制。方法:体外培养人胶质瘤U251细胞株,采用Lipofectamine 2000转染试剂将miR-148b-3p模拟物或阴性对照转染至U251细胞,分别记为miR-148b-3p组和阴性对照组,同时设置空白对照组。实时荧光定量聚合酶链反应(qRT-PCR)检测转染效果,Transwell实验检测各组U251细胞的侵袭能力,划痕实验检测各组U251细胞的迁移能力,酶联免疫吸附试验检测细胞上清液中基质金属蛋白酶2(MMP-2)和MMP-9的水平,Western blot法检测细胞中Wnt信号通路相关蛋白的表达。结果:qRT-PCR显示,miR-148b-3p组U251细胞中miR-148b-3p的表达水平为2.45±0.25,高于阴性对照组(0.97±0.10)和空白对照组(1.00±0.11),差异均有统计学意义(均P<0.05)。Transwell实验显示,miR-148b-3p组侵袭细胞数为(50.62±5.36)个,与阴性对照组[(108.84±10.14)个]和空白对照组[(113.40±10.06)个]比较,差异均有统计学意义(均P<0.05)。划痕实验结果显示,miR-148b-3p组细胞的迁移率为(23.19±2.50)%,与阴性对照组[(51.81±5.25)%]和空白对照组[(52.06±5.33)%]比较,差异均有统计学意义(均P<0.05)。过表达miR-148b-3p能够抑制MMP-2和MMP-9的表达,下调Wnt1和GSK-3β蛋白的表达。结论:miR-148b-3p能够通过抑制Wnt信号通路的激活抑制人胶质瘤U251细胞的侵袭和迁移能力。Objective To investigate the effect of miR-148b-3p on invasion and migration of glioma cells and the possible molecular mechanism.Methods Human glioma U251 cells were cultures in vitro.MiR-148b-3p mimic or negative control was transfected into U251 cells by Lipofectamine 2000 transfection reagent,which were recorded as miR-148b-3p group and NC group,respectively.A blank control(Ctr group)was set.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the transfection effect.Transwell assay was used to detect the invasive ability of U251 cells in each group.The scratch test was used to detect the migration ability of U251 cells in each group.The concentrations of matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-9)in the supernatant of the cells were determined by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the expressions of Wnt signaling pathway-related proteins in cells.Results The qRT-PCR experiment showed that the expression level of miR-148b-3p in U251 cells of miR-148b-3p group(2.45±0.25)was significantly higher than that of NC group(0.97±0.10)and Ctr group(1.00±0.11)(P<0.05).Transwell and scratch experiments showed that the number of invasive cells(50.62±5.36)in miR-148b-3p group was significantly lower than those in NC group(108.84±10.14)and Ctr group(113.40±10.06)(P<0.05).The results of the scratch experiment showed that the cell migration rate of miR-148b-3p group(23.19±2.50)% was significantly lower than those of NC group(51.81±5.25)% and Ctr group(52.06±5.33)%(P<0.05).Overexpression of miR-148b-3p inhibited the expressions of MMP-2 and MMP-9,downregulated the expressions of Wnt1 and GSK-3β protein.Conclusion miR-148b-3p can inhibit the invasion and migration of human glioma U251 cells by inhibiting the activation of Wnt signaling pathway.

关 键 词：miR-148b-3p WNT信号通路 胶质瘤细胞 侵袭 迁移 

分 类 号：R739.4[医药卫生—肿瘤]