微小RNA miR-148b-3p在人胶质瘤细胞中的表达及意义

The expression and significance of miR-148b-3p in human glioma cells

目的检测微小RNA(microRNA,miR)-148b-3p在人胶质瘤细胞中的表达水平,并探讨其对胶质瘤细胞增殖和侵袭的影响。方法荧光定量逆转录聚合酶链反应(qRT-PCR)方法检测胶质瘤细胞A172和正常星形胶质细胞HA1800中miR-148b-3p的表达水平。合成miR-148b-3p mimics及阴性对照(NC),用Lipofectamine 2000将miR-148b-3p mimics和NC转染至人胶质瘤细胞A172。分别采用噻唑蓝(MTT)法、Transwell实验、流式细胞术检测转染miR-148b-3p mimics对胶质瘤细胞A172增殖、侵袭及细胞周期的影响。结果 miR-148b-3p在胶质瘤细胞A172中的相对表达量为0.27±0.06,显著低于在正常星形胶质细胞HA1800中的相对表达量1.16±0.21(P<0.01)。MTT检测到miR-148b-3p mimics组A172细胞增殖率在转染后显著降低,转染24h、48h及72h后,细胞增殖率分别下降(12.33±0.46)%、(18.35±0.64)%和(30.54±1.22)%。Transwell实验结果显示miR-148b-3p mimics组A172细胞穿膜细胞数为23.1±1.8,显著低于NC组的87.1±4.6(P<0.01),miR-148b-3p mimics能够抑制A172细胞的侵袭能力。流式细胞术结果显示miR-148b-3p mimics显著阻滞A172细胞从G0/G1期向S期转换。结论 miR-148b-3p在胶质瘤细胞中低表达,miR-148b-3p在胶质瘤细胞中发挥抑制肿瘤增殖和侵袭的作用。

Objective To explore the expression of miR-148 b-3 p in human glioma cells and the influence of miR-148 b-3 p on proliferation and invasion of glioma cells.Methods The expressions of miR-148 b-3 p were detected by quantitative reverse transcriptase-polymerase chain reaction（qRT-PCR）in human glioma cells A172 and human astrocytes HA1800.miR-148 b-3 p mimic and miR-148 b-3 p NC were designed and transfected into glioma cell line A172 using Lipofectamine 2000.The effects of miR-148 b-3 p mimic on proliferation,invasion and cell cycle were analyzed by MTT assay,transwell assay and flow cytometry,respectively.Results The expression level of miR-148 b-3 p was 1.16±0.21 in HA1800 and 0.27±0.06 in A172 cells.The expression level of miR-148 b-3 p was significantly decreased in A172 cells compared with HA1800 cells（P〈0.01）.MTT results showed that the viability of A172 cell was significantly inhibited by transfection with miR-148 b-3 p mimic.24,48 and 72 hafter miR-148 b-3 p mimic transfection,cell proliferation rate decreased by（12.33±0.46）%,（18.35±0.64）%and（30.54±1.22）%,respectively.The in vitro transwell assay revealed that the number of invasive cells in miR-148 b-3 p mimic group was significantly decreased than in the control group（P〈0.01）.The invasiveness of A172 cells was significantly suppressed after transfected with miR-148 b-3 p mimic.Flow cytometry results showed that the cell population in the G1 phase was increased but the S phase population was decreased in miR-148 b-3 p mimic transfection group.Conclusion The expression level of miR-148 b-3 p was significantly decreased in glioma cell line.And miR-148 b-3 p plays a key regulatory role in the proliferation and invasion of human glioma cells.