胰岛素样生长因子结合蛋白2促进神经胶质瘤干细胞的增殖与迁移侵袭

Insulin-like growth factor binding protein-2 promotes the proliferation, migration and invasion of glioma stem cells

背景:目前关于血清胰岛素样生长因子结合蛋白2(recombinant insulin like growth factor binding protein 2,IGFBP-2)对神经胶质瘤影响的分子机制仍不明确,并且内源性IGFBP-2无法解释血清中IGFBP-2的一系列作用,需进一步阐明外源性IGFBP-2的作用。目的:观察IGFBP-2干预后神经胶质瘤干细胞的增殖与迁移侵袭能力变化。方法:采用免疫磁珠技术从人胶质瘤细胞U251细胞中分离CD133^+胶质瘤干细胞,免疫荧光染色检测细胞CD133、Nestin的表达进行鉴定;将CD133^+胶质瘤干细胞分2组干预,以500μg/L IGFBP-2干预48 h作为实验组,未经IGFBP-2干预作为对照组。CCK-8法检测细胞增殖能力,Transwell小室实验检测细胞迁移、侵袭能力,采用Western blot检测PCNA、Ki-67、ERK及p-ERK蛋白表达。结果与结论:(1)实验组培养2-5 d的吸光度值明显高于对照组,差异有显著性意义(P<0.05);实验组细胞增殖相关蛋白PCNA、Ki-67表达高于对照组,差异有显著性意义(P<0.05);(2)实验组迁移细胞数明显多于对照组,差异有显著性意义(P<0.05);(3)实验组侵袭细胞数明显多于对照组,差异有显著性意义(P<0.05);(4)两组ERK蛋白表达无差异,但实验组p-ERK蛋白明显高于对照组,差异有显著性意义(P<0.05);(5)结果表明,IGFBP-2可促进神经胶质瘤干细胞的增殖、迁移与侵袭,并且ERK通路可能在其中发挥了一定的作用。

BACKGROUND:To date,the molecular mechanism of recombinant insulin-like growth factor binding protein 2(IGFBP-2)on glioma is still unclear.Endogenous IGFBP-2 cannot explain a series of roles of IGFBP-2 in serum,and the roles of exogenous IGFBP-2 should be further elucidated.OBJECTIVE:To observe the effect of IGFBP-2 on the proliferation,migration and invasion of glioma stem cells.METHODS:CD133+glioma stem cells were isolated from U251 human glioma cells by immunomagnetic bead technique.CD133 and nestin expression was detected by immunofluorescence staining.CD133+glioma stem cells were divided into two groups:the cells were treated with 500μg/L IGFBP-2 for 48 hours as experimental group,and the cells without intervention of IGFBP-2 as control group.Cell counting kit-8 method was used to detect cell proliferation ability.Transwell chamber assay was used to detect cell migration and invasion ability.Western blot assay was used to detect the expression of PCNA,Aki67,ERK and p-ERK proteins.RESULTS AND CONCLUSION:The cell absorbance of the experimental group was significantly higher than that of the control group at 2-5 days of culture(P<0.05),and the expression of PCNA and Ki-67 in the experimental group was also significantly higher than that in the control group(P<0.05).The number of migrated cells and invasive cells in the experimental group was significantly higher than that in the control group(P<0.05).Although there was no difference in the expression of ERK protein between the two groups,the expression of p-ERK protein in the experimental group was significantly higher than that in the control group(P<0.05).To conclude,IGFBP-2 can promote the proliferation,migration and invasion of glioma stem cells,in which the ERK pathway may play a certain role.