胶质瘤干祖细胞SU2分化抑制、侵袭性与miRNA-125b和MMP-9关系的研究

Study of relationships between the differential inhibition and invasiveness of glioma stem-progenitor cells SU2 and the expressions of miRNA-125b and MMP-9

目的探讨胶质瘤干祖细胞SU2分化抑制、侵袭性与基质金属蛋白酶(MMP-9)和miRNA-125b的关系。方法胶质瘤干祖细胞SU2经丙戊酸钠(sodium valproate,VPA)诱导分化得到SU2分化细胞。U87胶质瘤细胞经无血清、含生长因子的培养基培养得到U87胶质瘤干祖细胞。免疫荧光双标记法检测SU2及其分化细胞中CD133、Nestin、胶质纤维酸性蛋白(GFAP)、β微管蛋白(β-tubulinⅢ)的表达。Transwell实验检测SU2和U87胶质瘤干祖细胞的侵袭情况。实时荧光定量PCR检测SU2及U87胶质瘤干祖细胞中miRNA-125b和MMP-9基因表达。结果 SU2及其分化细胞均表达β-tubulinⅢ和GFAP。SU2细胞侵袭数明显多于U87细胞(P=0.026 1)。SU2细胞中的miRNA-125b和MMP-9表达明显高于U87细胞(均P<0.05)。结论胶质瘤干祖细胞SU2分化抑制和高侵袭的生物学特征与miRNA-125b和MMP-9的高表达有关,MMP-9可作为miRNA-125b调控的靶基因进一步研究。

Objective To explore the relationships between the differential inhibition,invasiveness of glioma stem-progenitor cells SU2 and the expressions of matrix metalloproteinases 9（MMP-9） and miRNA-125b.Methods The glioma stem-progenitor cells SU2 were induced into differentiated SU2 cells by sodium valproate（VPA）.U87 cells were induced into U87 stem-progenitor cells by using culture medium with growth factors but without FBS.The expressions of differentiated cell markers including CD133,Nestin,β-tubulin Ⅲ and glial fibrillary acidic protein（GFAP） in SU2 and differentiated SU2 cells were tested by double immunofluorescence method.The invasiveness of SU2 and U87 stem-progenitor cells was tested by Transwell invasion assay.The gene expressions of miRNA-125b and MMP-9 in SU2 and U87 stem-progenitor cells were detected by real-time fluorescent quantitative PCR.Results SU2 and differentiated SU2 cells were positive for β-tubulin Ⅲ and GFAP.The invasive cells of SU2 cells were more than those of U87 cells（P = 0.026 1）.The gene expressions of miRNA-125b and MMP-9 were higher in SU2 cells than in U87 cells（both P 0.05）.Conclusions Differential inhibition and high invasive ability of SU2 may be related to the high expression of miRNA-125b and MMP-9.MMP-9 can be further studied as a target gene of miRNA-125b.