TCLA诱导大鼠脑胶质瘤细胞增殖抑制及凋亡的研究  

Apoptosis and Differentiation Induced with TCLA in Rat Glioblastoma Cells in vitro

作　　者：扈及雷[1] 王修杰[2] 周培志[1] 王成德[1] 徐建国[1] 姜曙[1] 

机构地区：[1]四川大学华西医院神经外科,成都610041 [2]四川大学生物治疗国家重点实验室实验肿瘤研究室

出　　处：《四川大学学报（医学版）》2008年第4期570-574,共5页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅科技支撑项目(批准号2008SZ0033)资助

摘　　要：目的探讨共轭三烯酸(TCLA)对大鼠脑胶质瘤细胞体外增殖抑制、凋亡作用及其作用机制。方法采用细胞增殖实验(MTT法)、克隆形成实验、BrdU掺入实验检测TCLA对大鼠脑胶质瘤细胞系C6的体外增殖抑制作用;Hoechst33342染色、流式细胞术检测细胞凋亡和细胞周期分布;RT-PCR检测调亡相关基因腺苷二磷酸核糖转移酶1(ADPRTL1)、细胞色素P4501A1(CYP1A1),过氧化物酶增殖体激活受体-γ(PPAR-γ)的表达。结果MTT法发现,TCLA可显著抑制C6细胞株增殖(40μmol/L,72h,细胞存活率56.71%±0.98%),并存在量-效、时-效关系;克隆形成下降,40μmol/L的TCLA可完全抑制克隆形成;实验组BrdU标记率63.1%±1.0%,较对照组(95.6%±1.4%)明显降低(P<0.05);Hoechst33342荧光染色示,实验组凋亡率10.0%±0.3%,较对照组(1.6%±0.6%)明显升高(P<0.05);流式细胞术分析结果示,TCLA使C6细胞凋亡指数升高,G0/G1期细胞比例增加,S期细胞比例减少,与对照组的差异均有统计学意义(P<0.05);RT-PCR结果显示,实验组ADPRTL1、CYP1A1、PPAR-γmRNA表达均较对照组增强,差异均具有统计学意义(P<0.05)。结论TCLA对大鼠脑胶质瘤细胞具有抑制增殖和诱导凋亡作用。其作用机制可能与抑制肿瘤细胞DNA合成、细胞周期阻滞、上调凋亡相关基因(ADPRTL1、CYP1A1、PPAR-γ)表达有关。Objective To investigate the effect of conjugated trienoic fatty acids on the growth inhibition,apoptosis in rat glioblastoma cells and to elucidate its mechanism of activity.Methods Rat glioblastoma cells were tested in vitro cytotoxicity,colony formation inhibition,Brdu incorporation after treatment with TCLA.Its effect of apoptosis induction was detected through Hoechst33342 staining,cell cycle analysis.The expressions of ADPRTL1,CYP1A1 and PPAR-γ genes were detected through RT-PCR.Results After TCLA treatment,the proliferation of C6 cells were inhibited（40 μmmol/L,72 h,viability 56.71%±0.98%）,this action acted as dose-time-dependent relations;colony formation decreased significantly（40 μmmol/L,0）and BrdU labeling index of cancer cells decreased（63.1%±1.0% vs 95.6%±1.4%）;apoptotic cells increased;By FCM analysis,the apoptotic indices increased,the cells increased in G0/G1 phase,decreased in S phase,which have signigicant difference;RT-RCR showed that TCLA signigicantly increased the level of ADPRTL1,CYP1A1,and PPAR-γ mRNA expression.Conclusion The findings in this experimental study suggested that TCLA has potent cytotoxicity and induction apoptosis in human and rat glioblastoma cells,its mechanism of activity might be associated with the inhibition of DNA synthesis,cell cycle arrest,up-regulation the expression of apoptosis related genes ADPRTL,CYP1A1,PPAR-γ.

关 键 词：共轭三烯酸 胶质瘤 增殖抑制 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]