Shang Chao, Guo Yan, Hong Yang, Liu Yun-hui, Xue Yi-xue
Department of Neurobiology, China Medical University, No. 92 Beier Road, Heping District, Shenyang, 110001, People's Republic of China.
Mol Biol Rep. 2015 Mar;42(3):721-7. doi: 10.1007/s11033-014-3820-3. Epub 2014 Nov 14.
To investigate whether miR-21 can affect the apoptosis and proliferation of glioblastoma cancer stem cells (GSCs) from down-regulating FASLG. The expression of miRNA-21 was detected by quantitative real-time PCR in normal brain tissue and glioblastoma samples, and the changes of miRNA-21 expression between GSCs and non-GSCs were also detected. The apoptosis and proliferation ability of miR-21 in GSCs were analyzed by MTT and flow cytometry assay after anti-miR-21 transfection. For the regulation mechanism analysis of miR-21, TargetScan, PicTar and microRNA were selected to predict some potential target genes of miR-21. The predicted gene was identified to be the direct and specific target gene of miR-21 by luciferase activities assay and western blot. RNA interference technology was used to confirm the apoptosis and proliferation effects of miR-21 were directly induced by FASLG. The expression of miR-21 increased significantly in glioblastoma contrast to normal brain tissue, and miR-21 up-regulated in GSCs remarkably. The proliferation of GSCs cell could be inhibited with high-expression of miR-21 and this effect could be restored by miR-21 knocked down. Mechanism analysis revealed that FASLG was a specific and direct target gene of miR-21. The advanced effects of anti-miR-21 on GSCs apoptosis and proliferation were mediated by expression of silenced FASLG. In summary, aberrantly expressed miR-21 regulates GSCs apoptosis and proliferation partly through directly down-regulating FASLG protein expression in GSCs and this might offer a new potential therapeutic stratagem for glioblastoma.
为研究miR-21是否可通过下调FASLG影响胶质母细胞瘤癌干细胞(GSCs)的凋亡和增殖。采用定量实时PCR检测正常脑组织和胶质母细胞瘤样本中miRNA-21的表达,并检测GSCs和非GSCs之间miRNA-21表达的变化。抗miR-21转染后,通过MTT和流式细胞术分析miR-21对GSCs凋亡和增殖能力的影响。为分析miR-21的调控机制,选择TargetScan、PicTar和microRNA预测miR-21的一些潜在靶基因。通过荧光素酶活性测定和蛋白质印迹法鉴定预测基因是miR-21的直接和特异性靶基因。采用RNA干扰技术证实miR-21的凋亡和增殖作用是由FASLG直接诱导的。与正常脑组织相比,胶质母细胞瘤中miR-21的表达显著增加,且GSCs中miR-21明显上调。miR-21高表达可抑制GSCs细胞增殖,敲低miR-21可恢复此效应。机制分析显示FASLG是miR-21的特异性直接靶基因。抗miR-21对GSCs凋亡和增殖的促进作用是通过沉默FASLG的表达介导的。综上所述,异常表达的miR-21部分通过直接下调GSCs中FASLG蛋白表达来调节GSCs的凋亡和增殖,这可能为胶质母细胞瘤提供一种新的潜在治疗策略。