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目的研究三氧化二砷(As2O3)在体外是否具有抗鼠胶质瘤作用,并对其作用机制进行初步探讨。方法应用不同浓度As2O3,且在不同时间点分别处理C6胶质瘤细胞株及原代培养的正常鼠神经胶质细胞,采用甲基噻唑基四唑(MTT)法,观察As2O3对C6胶质瘤细胞株及正常鼠神经胶质细胞生长的影响,以透射电镜、Hoechst33342和碘化丙啶(PI)双重荧光染色检测两种细胞凋亡的形态变化,并用异硫氰酸荧光素(FITC)标记的Annexin-V和PI双标记法通过流式细胞仪检测细胞凋亡率。结果MTT法发现,As2O30.5~8.0μmol/L的浓度均可显著抑制C6胶质瘤细胞株的生长,而对正常原代神经胶质细胞株的抑制作用较弱。经As2O3作用后,透射电镜、Hoechst33342/PI双染荧光均观察到C6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As2O3浓度的增大和时间的延长,C6胶质瘤细胞株的凋亡率明显上升,而正常神经胶质细胞的凋亡率要明显小于C6胶质瘤细胞株。结论As2O3在体外可显著抑制C6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As2O3作用的时间和剂量的增加而增加。但As2O3对正常神经胶质细胞生长的抑制作用较弱,提示As2O3抑制细胞生长的作用具有一定的选择性。
Objective To study whether arsenic trioxide (As2O3) has an anti-mouse glioma effect in vitro and to explore its mechanism. Methods As2O3 was treated with different concentrations of As2O3, and C6 glioma cell lines and primary cultured normal rat glial cells were treated at different time points. Methyl thiazolyl tetrazolium (MTT) method was used to observe the effects of As2O3 on C6 glioma Cell lines and normal rat glial cells. The morphological changes of apoptosis were detected by transmission electron microscopy, Hoechst33342 and PI staining, and labeled with fluorescein isothiocyanate (FITC) Annexin-V and PI double labeling method was used to detect the apoptosis rate by flow cytometry. Results MTT assay showed that the concentration of As2O30.5 ~ 8.0μmol / L can significantly inhibit the growth of C6 glioma cell lines, while the inhibition of normal primary glial cell lines is weak. After treated with As2O3, the apoptotic morphological changes of C6 glioma cells were observed by transmission electron microscopy and Hoechst33342 / PI double staining. Flow cytometry was performed using Annexin-V-FITC / PI double labeling assay. With the increase of As2O3 concentration and time, the apoptosis rate of C6 glioma cell line increased obviously, while the apoptosis rate of normal glial cells was significantly smaller than C6 glioma cell line. Conclusion As2O3 can significantly inhibit the growth of C6 glioma cell lines in vitro, and its mechanism is related to the induction of tumor cell apoptosis, and the apoptosis rate increases with the time and dose of As2O3. However, the inhibitory effect of As2O3 on normal glial cell growth was weak, suggesting that As2O3 could inhibit the cell growth with a certain selectivity.