三氧化二砷联合顺铂抑制C6胶质瘤细胞增殖的实验研究  

Effect of arsenic trioxide in combination with cisplatin on the proliferation of glioma C6 cells

作　　者：王志刚[1] 钟根深[1] 马鹏举[1] 岳双柱[1] 周国胜[1] 

机构地区：[1]新乡医学院第一附属医院神经外科/河南省神经病学研究所,河南卫辉453100

出　　处：《中国医药生物技术》2014年第3期167-173,共7页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金青年科学基金(81201765);河南省卫生科技创新型人才工程(4160)

摘　　要：目的探讨三氧化二砷(As2O3)联合顺铂(CDDP)抑制C6神经胶质瘤细胞增殖情况及其分子机制。方法体外实验采用MTT法检测对C6神经胶质瘤细胞的抑制率,流式细胞法检测C6胶质瘤细胞的凋亡及周期,Western blot检测caspase-3、PTEN、PI3K、Akt蛋白表达水平;体内实验采用免疫组化法检测caspase-3、IκB-α调控因子的表达,并观察大鼠的生存情况。结果 As2O3、CDDP及联合应用分别作用于C6神经胶质瘤细胞24 h后的细胞增殖最大抑制率分别为34.37%、31.73%和76.40%,其相应药物浓度分别为12.5、10.0、(12.5+10.0)μmol/L;两者联用后对C6胶质瘤细胞作用24 h后的凋亡率分别为从单独用药时的(4.60±1.12)%、(14.59±0.79)%提高到(36.13±1.55)%;采用PI单染检测的细胞周期也进一步证明了联合用药增加了C6细胞的周期抑制作用。Caspase-3、PTEN蛋白在联合组C6胶质瘤细胞中表达显著上调,而PI3K、Akt蛋白在联合组C6胶质瘤细胞中表达下调;免疫组化法检测结果显示,caspase-3调控因子在联合组用药的C6胶质瘤细胞中的表达上调,而IκB-α调控因子表达下调;但体内实验表明,在18 d观察期内各组大鼠的生存期无明显差异(P=0.2531)。结论 As2O3联合CDDP通过上调caspase-3和PTEN,下调PI3K、Akt及IκB-α抑制胶质瘤细胞体内外的生长。Objective To investigate the effects of arsenic trioxide in combination with cisplatin on the proliferation of glioma cells C6. Methods In the in vitro experiment, the proliferation inhibition of glioma cell line C6 was assessed by MTT assay, and the cell cycle distribution and apoptosis ratio of the cells were detected by flow cytometry. The expression levels of caspase-3, PTEN, PI3K and Akt were examined by Western blot. In the in vivo experiment, the expression levels of caspase-3 and IκB-α were measured by immunohistochemical methods, and the rats’ life span was observed. Results At the dosage of 12.5 μmol/L of As2O3 and 10.0 μmol/L of CDDP, the proliferation inhibitory ratios of As2O3, CDDP and＆amp;nbsp;co-treatment group on glioma cell line C6 after 24 hours were 34.37%, 31.73% and 76.40%, respectively. Correspondingly, the apoptosis ratios after 24 hours were （4.60 ± 1.12）%, （14.59 ± 0.79）%and （36.13 ± 1.55）%respectively. Apoptosis induction was also proved by FACS using PI-stained method. The expression levels of caspase-3 and PTEN were up-regulated, while the expression levels of PI3K and Akt were down-regulated in the co-treatment group. For in vivo experiment, the expression of caspase-3 was up-regulated and IκB-αwas down-regulated in the co-treatment group. However, the rats’ survival rate in different treatment groups was similar in the limited observation time （P=0.2531）. Conclusion As2O3 combined with CDDP has potent inhibitory action on glioma cells proliferation through up-regulating caspase-3 and PTEN and down-regulating PI3K, Akt and IκB-αexpression levels.

关 键 词：顺铂 神经胶质瘤 细胞凋亡 三氧化二砷 药物疗法 联合 

分 类 号：R739.4[医药卫生—肿瘤]