雄黄体外诱导C6胶质瘤细胞凋亡的特点及分析  

Cellular apoptosis of C6 glioma induced by realgar in vitro

作　　者：王汉斌[1,2] 庞琦[2] 富壮[1] 赵光宇[2] 丁锋[2] 栾立明[2] 郭华[2] 许家军[2] 许尚臣[2] 

机构地区：[1]北京积水潭医院神经外科,北京100035 [2]山东大学附属省立医院神经外科,济南250021

出　　处：《山东大学学报（医学版）》2010年第1期21-24,29,共5页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金(Y2005C31);山东省医药卫生科技发展计划(2007HZ064);山东省科学技术发展计划(2007GG30002010)资助

摘　　要：目的观察雄黄对体外培养的C6胶质瘤细胞的生长抑制作用,探讨雄黄诱导的细胞凋亡特点和可能机制。方法以雄黄溶液对体外培养的大鼠C6胶质瘤细胞进行诱导,通过MTT法观察浓度和时间对细胞存活率的影响,通过光镜、电镜观察细胞的抑制情况;采用AnnexinV-FITC-PI双染流式细胞术观察药物引起的细胞凋亡情况,采用RT-PCR法观察诱导过程中Bcl-2、Bax和c-myc3个基因的表达情况。结果MTT结果显示,不同浓度的雄黄溶液可引起细胞不同程度的抑制;光镜及透射电镜下可观察到药物引起的细胞凋亡变化。Annexin V-FITC-PI双染显示,细胞在25mg/L雄黄作用24、48h后的凋亡率分别为(10.13±3.14)%、(41.04±17.44)%,与对照组相比差异有统计学意义;RT-PCR观察到Bax基因表达的升高和Bcl-2、c-myc基因表达降低。结论雄黄对C6细胞的抑制具有时间和浓度依赖性,浓度越高,越明显,此抑制与雄黄引起的凋亡有关,凋亡过程中伴有Bax基因表达的增加和Bcl-2、c-myc的降低。Objective To investigate the effects of realgar on C6 glioma and its possible mechanisms concerning cell apoptosis in vitro. Methods Rat C6 glioma cells were cultured under standard conditions. Realgar solution was used at several concentrations on glioma cells for 24 h and 48 h. The inhibitory effect was observed by MTT,microscopy and TEM. FCM was used to detect the apoptosis of glioma cells after Annexin V-FITC-II double staining,and RT-PCT was used for the detection of the three apoptosis genes. Results The MTT method showed gradient cell growth inhabitation at gradient concentrations. Cell apoptosis was observed via microscope and TEM. Annexin V-FITC-PI double staining showed the apoptosis rate of realgar at 25 mg/L after 24 h and 48 h was （10.13±3.14）% and （41.04±17.44）% respectively,which was significantly different from that of the control group. The up-regulation of Bax mRNA and down-regulation of Bcl-2 and c-myc mRNA were also observed by RT-PCR. Conclusion The effects of realgar on C6 glioma are positively time-and concentration-related. This phenomenon is related to apoptosis induced by realgar. The up-regulation of Bax and down-regulation of Bcl-2 and c-myc maybe play important roles in the process.

关 键 词：雄黄 神经胶质瘤 细胞凋亡 

分 类 号：R739.4[医药卫生—肿瘤]