胶质瘤干细胞分离鉴定及其化疗敏感性研究

Isolation and identification of brain glioma stem cell and its chemotherapeutic sensitivity

目的:探讨胶质瘤干细胞的分离、培养、鉴定及其与胶质瘤细胞化疗敏感性方面的差异。方法:将3株人胶质瘤细胞SHG-44、U251、U87MG分别接种于无血清培养基,从中分离出各自胶质瘤干细胞;扫描电镜观察胶质瘤干细胞的形态;RT-PCR法检测胶质瘤干细胞CD133、GFAP的表达;CCK8法检测化疗药物顺铂及长春新碱分别作用后胶质瘤细胞及其干细胞活性的变化。结果:从3株胶质瘤细胞中分离出的胶质瘤干细胞,可在体外增殖及传代,表达特异性标志物CD133,不表达GFAP。CCK8法检测出胶质瘤干细胞较胶质瘤细胞更为耐药(均P<0.05)。结论:胶质瘤干细胞较胶质瘤细胞化疗敏感性降低。

Objective： To explore the methods of isolation, culture and identification of glioma stem cells（ GSCs）, and to study the difference of chemotherapeutic sensitivity between the GSCs and the glioma cells. Methods： Three strains of glioma cells were respectively seeded into serum-free medium and then the GSCs of each were isolated. The morphology of GSCs was observed under the scanning electron microscope. The expression of CD133 and GFAP in GSCs was detected by RT-PCR. The CCK8 was used to evaluate the difference of proliferation between the glioma cells and GSCs after they were interved with cisplatin and vincristine respectively. Results： The GSCs isolated from glioma cells had the ability to proliferate and renew in vitro. It showed that there was expression of CD133 and no expression of GFAP in GSCs. Compared with glioma cells, GSCs were more resistant to drug therapy （all P 〈 0.05）. Conclusion： GSCs are more resistant to chemotherapy than glioma cells.