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目的:研究卡氮芥(BCNU)是否具有抑制鼠胶质瘤的作用,并对其作用机制进行初步探讨。方法:应用不同浓度BCNU,且在不同时间点分别处理9L胶质瘤细胞株,采用四甲基噻唑蓝比色(MTT)法,观察BCNU对9L胶质瘤细胞株生长的影响,以透射电镜、Hoechst33342/PI双染荧光和TUNEL检测细胞凋亡的形态变化,并用Annexin-v-FITC/PI双标记法流式细胞仪检测细胞凋亡率。结果:MTT法发现,BCNU0.003～3000μmol.L-1的浓度均可显著抑制9L胶质瘤细胞株的生长。经BCNU作用后,透射电镜、Hoechst33342/PI双染荧光和TUNEL均观察到9L胶质瘤细胞发生了显著的凋亡形态改变;运用Annexin-V-FITC/PI双标记法在流式细胞仪检测显示,随BCNU浓度的增大和时间的延长,9L胶质瘤细胞株的凋亡率明显上升。结论:BCNU在体外可显著抑制9L胶质瘤细胞株生长,其机制与诱导肿瘤细胞凋亡有关,且凋亡率随BCNU作用的时间和剂量的增加而增加。
Objective: To study whether Carnitine (BCNU) has the effect of inhibiting glioma and to explore its mechanism. Methods: Different concentrations of BCNU were applied and 9L glioma cell lines were treated at different time points. The effect of BCNU on the growth of 9L glioma cell line was observed by MTT method. Transmission electron microscopy , Hoechst33342 / PI double staining and TUNEL. The apoptotic rate was detected by Annexin-v-FITC / PI double-labeled flow cytometry. Results: MTT assay showed that the concentration of BCNU0.003 ~ 3000μmol.L-1 significantly inhibited the growth of 9L glioma cell lines. After BCNU, the apoptosis of 9L glioma cells was observed by transmission electron microscopy, Hoechst33342 / PI double staining and TUNEL staining. Annexin-V-FITC / PI double labeling method was used to detect the apoptosis in 9L glioma cells by flow cytometry The results showed that with the increase of BCNU concentration and the prolongation of time, the apoptosis rate of 9L glioma cell line increased obviously. CONCLUSION: BCNU can significantly inhibit the growth of 9L glioma cell lines in vitro. The mechanism is related to the induction of tumor cell apoptosis, and the apoptosis rate increases with the time and dose of BCNU.