【摘要】 目的:探讨miR-183对胶质母细胞瘤放射抵抗等恶性行为的调控作用及其机制。方法:利用实时定量聚合酶链反应(RT-qPCR)方法检测15例胶质母细胞瘤及瘤周正常脑组织及4株胶质母细胞瘤细胞株(U87、U251、M059J、M059K)中miR-183表达水平;采用阳离子脂质体法瞬时转染构建miR-183过表达的细胞体系;应用MTT细胞增殖实验检测细胞增殖能力;利用平板克隆形成实验检测辐射敏感性;采用划痕实验和Transwell实验检测miR-183对胶质母细胞瘤细胞侵袭及迁移能力的调控作用;利用蛋白印迹(Western blot)实验检测miR-183对上皮间质转化(EMT)相关基因表达水平的调控作用。结果 :miR-183在胶质母细胞瘤及胶质母细胞瘤细胞株中低表达。随后通过构建过表达miR-183的细胞系,发现miR-183高表达状态的胶质母细胞瘤细胞对射线的敏感程度增加,增殖能力、迁移能力和侵袭能力都有所下降,而且上调了上皮细胞表型相关E-钙黏蛋白的表达,下调了间质细胞表型相关波形蛋白的表达。结论:miR-183可能通过抑制EMT过程从而影响胶质母细胞瘤的恶性行为。更多还原

【Abstract】 Objective To investigate the regulation of miR-183 in inhibiting malignant behaviors such as radiation resistance in glioblastoma. Methods Using real-time quantitative polymerase chain reaction(RTq PCR) to detect miR-183 expression level in 15 glioblastoma tumor tissues with surrounding brain tissues and 4 glioblastoma cell lines(U87, U251, M059 J, M059K). Cationic liposomes transient transfection was used to construct cells which overexpressed miR-183. Cell proliferation ability was tested with MTT cell proliferation experiment. Clone formation experiment was conducted to detect radiation sensitivity. Wo-und scratch and Transwell experiment was used to detect invasion and migration ability that regulated by miR-183 in glioblastoma cells. Relevance between miR-183 and epithelial mesenchymal transformation(EMT) was examined by related protein expression with western blot. Results There was a low expression of miR-183 in glioblastoma tissue and cell lines. Radiation sensitivity increased in miR-183 overexpressed glioblastoma cells, while the proliferation,migration and invasion ability decreased. Overexpression of miR-183 also raised the expression of protein Ecadherin,but decreased the expression of vimentin, which indicated Mesenchymal Epithelial transformation.Conclusion MiR-183 could inhibit the EMT process to inhibit the malignant behavior of glioblastoma.更多还原