Wan Yi, Fei Xi-Feng, Wang Zhi-Min, Jiang Dong-Yi, Chen Han-Chun, Yang Jian, Shi Lei, Huang Qiang
Suzhou Kowloon Hospital, Shanghai Jiaotong University School of Medicine, Suzhou, Jiangsu 215021, PR China.
Chin J Cancer. 2012 Apr;31(4):207-14. doi: 10.5732/cjc.011.10336. Epub 2012 Feb 24.
MicroRNA (miR)-125b has been shown to play a potential role in the development of glioma stem cells. However, the relationship between miRNA and glioma stem cells is still elusive. This study was designed to elucidate this potential relationship. We established a highly invasive glioma stem cell and progenitor (GSCP) cell line SU3. SU3 cell suspensions were injected into nude mice brains in situ, and the invasiveness of graft tumors was analyzed using hematoxylin and eosin staining as well as immunohistochemistry. Real-time polymerase chain reaction (PCR) was used to measure the expression levels of miR-125b in SU3 and other cells. In vitro, SU3 cells expressed CD133 and nestin as well as differentiation markers glial fibrillary acidic protein (GFAP) and β-tubulin III, which were consistent with the characteristics of glioma stem cells. Scratch assays indicated that the migration ability of SU3 cells was stronger than that of U251 stem cells (U251s). In vivo, SU3 cells invaded into each part of the mouse brain from the caudate nucleus in a diffuse pattern and highly expressed invasive and proliferative cell markers matrix metalloprotease 2 (MMP2), MMP9, and Ki-67. Real-time PCR results revealed that the levels of miR-125b and MMP9 were significantly higher in SU3 and SU2, also a highly invasive GSCP cell line we established before, than in U251s. High expression of miR-125b both in newly established GSCPs, SU3, and long-term cultured GSCPs, SU2 suggests that miR-125b exhibits oncogene-like behavior. This behavior should be considered in further studies of miR-125b in cancer stem cells. Furthermore, MMP9, which plays a role in cancer stem cell invasion, may be a target gene of miR-125b.
微小RNA(miR)-125b已被证明在胶质瘤干细胞的发展中发挥潜在作用。然而,微小RNA与胶质瘤干细胞之间的关系仍不清楚。本研究旨在阐明这种潜在关系。我们建立了一种高侵袭性的胶质瘤干细胞和祖细胞(GSCP)系SU3。将SU3细胞悬液原位注射到裸鼠脑内,并用苏木精和伊红染色以及免疫组织化学分析移植瘤的侵袭性。采用实时聚合酶链反应(PCR)检测SU3和其他细胞中miR-125b的表达水平。在体外,SU3细胞表达CD133和巢蛋白以及分化标志物胶质纤维酸性蛋白(GFAP)和β-微管蛋白III,这与胶质瘤干细胞的特征一致。划痕试验表明,SU3细胞的迁移能力强于U251干细胞(U251s)。在体内,SU3细胞以弥漫性模式从尾状核侵入小鼠脑的各个部位,并高表达侵袭性和增殖性细胞标志物基质金属蛋白酶2(MMP2)、MMP9和Ki-67。实时PCR结果显示,SU3和SU2(我们之前建立的另一种高侵袭性GSCP细胞系)中miR-125b和MMP9的水平显著高于U251s。新建立的GSCPs SU3和长期培养的GSCPs SU2中miR-125b的高表达表明miR-125b表现出癌基因样行为。在进一步研究miR-125b在癌症干细胞中的作用时应考虑这种行为。此外,在癌症干细胞侵袭中起作用的MMP9可能是miR-125b的靶基因。