原发耐药胶质母细胞瘤细胞株的建立

Establishment of temozolomide-resistant cell line of primary human glioblastoma

目的通过原代培养胶质母细胞瘤(GBM)和体外替莫唑胺敏感性实验,建立原发耐药胶质母细胞瘤细胞模型。方法对术后病理确诊为GBM的3例脑组织通过酶消化法行原代培养,分别为GBM1、GBM2和GBM3;并与U87细胞进行对比。绘制4株细胞生长曲线,流式细胞仪分析4株细胞的增殖周期。MTS法检测8个浓度替莫唑胺对4株细胞生长的抑制率,Annexin-V-FITC/PI(磷脂结合蛋白)检测替莫唑胺作用后细胞凋亡率。结果 4株细胞均具备肿瘤细胞的典型特点。不同浓度替莫唑胺对GBM3细胞抑制率明显低于其他3株细胞。GBM3细胞株的凋亡率明显低于其他3株细胞。结论本研究成功培育3株原代胶质瘤细胞株并成功建立替莫唑胺原发耐药细胞株1株。

Objective To build primary drag-resistant glioblastoma （GBM） model by primary cultures of GBM cells and in-vitro temozolomide （TMZ） susceptibility test. Methods The glioma cells from 3 GBM patients were GBM1, GBM2 and GBM3 respectively, primarily cultured by tissue piece culture method and compared with U87 glioma cell lines. The growth curves of four cells lines were drawn and their proliferative cycles were analyzed using a flow cytometer. MTS was applied to detect the sensitivity of the four cell lines to eight different concentrations of TMZ. The TMZ-related apoptosis rate was detected by Annexin-V-FITC/PI. Results All the four cell lines were of typical characteristics of tumor cells. MTS method showed that inhibition of GBM3 glioblastoma cell line growth by different concentrations of TMZ was less than the other 3 cell lines. The apoptosis rate was lower in GBM 3 than the other three cell lines. Conclusion In this study, three primary GBM cell lines are successfully cultured and a primary TMZ-resistant cell model is successfully established.