敲减长链非编码RNA ATB抑制胶质瘤细胞增殖、迁移和侵袭

摘要

Objective To investigate the effects of long non-coding RNA activated by TGF-β(ATB) on the proliferation, migration and invasion of human U87glioma cell line.Methods The expression of ATB in human U87 glioma cell line and human normal astrocyte(HEB) were detected by real-time quantitative PCR(qRT-PCR).And based on this, shRNA ATB plasmid were constructed ,which could knockdown ATB expression after transfected into U87 cells.The proliferation ability of cell was investigated through MTT assay and the colony-formation ability was tested by the cell cloning assay.Cell migration and invasion were measured by non-Matrigel transwell assay and Matrigel transwell assay in vitro.Results Using qRT-PCR analysis, we found ATB expression levels in U87 cell line was significantly increased compared with HEB(P<0.01).The expression of ATB was remarkable reduced after U87 transfected with shRNA-ATB plasmid(the experimental group) compared with U87 cells transfected with shRNA control(the control group).Cloning formation experiment indicated that colony-formation ability was decreased than that of control group(P<0.01).MTT assay indicated that the number of viable cell was lower than that of control group(P<0.01).In addition, Transwell assay further verified that knockdown of ATB significantly inhibited the migration and invasion ability of U87 cells(P<0.01).Conclusion ATB knockdown inhibit glioma cell proliferation, migration and invasion, which indicates that ATB may represent a potential therapeutic target for the treatment of human glioma.%目的 采用shRNA ATB敲减胶质瘤U87细胞中ATB后,研究其对人胶质瘤细胞增殖、迁移和侵袭的影响.方法 本研究采用实时定量PCR(qRT-PCR)方法检测ATB在人胶质瘤细胞系U87与正常人脑胶质细胞HEB中的表达情况.并在此基础上构建了ATB表达载体shRNA ATB 质粒,利用其转染人胶质瘤U87细胞株,获取低表达ATB的U87细胞.应用MTT法检测敲减ATB后对U87细胞增殖的影响;应用细胞克隆实验检测敲减ATB后对U87细胞克隆增殖能力的影响;应用Transwell实验检测敲减ATB后对U87细胞迁移和侵袭能力的影响.结果 qRT-PCR结果显示,与正常人脑胶质细胞HEB相比,人脑胶质瘤细胞系U87中ATB 表达明显上调(P<0.01);与shRNA对照组相比,转染了shRNA-ATB实验组能显著减少ATB 水平(P<0.01);细胞克隆实验显示shRNA-ATB实验组细胞克隆形成能力较shRNA对照组明显降低(P<0.01);MTT结果表明敲减细胞内ATB后细胞增殖的速率明显低于shRNA对照组(P<0.01).此外,Transwell实验进一步验证了敲减胶质瘤U87细胞中的ATB后,细胞迁移和侵袭能力明显受到抑制(P<0.01).结论 靶向敲减U87细胞中的ATB后能够抑制其增殖、迁移和侵袭,表明ATB基因可能是胶质瘤患者的潜在治疗靶点.