摘要
目的研究Hedgehog信号通路特异性阻断剂环靶明(cyclopamine)在体内及体外对人胶质母细胞瘤A172细胞生长的影响。方法用四甲基偶氮唑盐比色法(MTT)检测环靶明对A172细胞增殖的抑制作用,流式细胞术检测细胞凋亡率;反转录-聚合酶链反应(RT-PCR)检测环靶明处理前后Smoothened蛋白(Smoothened,SMO)、GLI家族锌指蛋白1(GLI-Kruppel family member 1,GLI1)在A172中的mRNA表达及变化,免疫印迹法(Western Blot)检测环靶明处理前后SMO、GLI1在A172中的蛋白表达及变化;环靶明对A172裸鼠皮下移植瘤生长的影响。结果环靶明抑制A172细胞的增殖,其作用呈剂量和时间依赖性;经不同浓度的环靶明作用48 h后A172细胞的凋亡率逐渐升高,明显高于对照组的凋亡率(P<0.01),并存剂量依赖性。SMO、GLI1的mRNA和蛋白均在A172中表达,环靶明下调A172的SMO、GLI1表达。环靶明可抑制A172裸鼠皮下移植瘤的生长。结论在体内及体外阻断Hedgehog信号通路可抑制人胶质母细胞瘤细胞A172的生长。
AbstractObjectiveThe study aims to investigate Hedgehog signaling pathway specific blocker （cyclopamine） effects human glioblastoma A172 cells growth in vivo and in vitro.MethodsThe proliferation of A172 cells was detected with Methylcyclopentadienyl Manganese Tricarbonyl （MTT） method. Cells apoptotic rate was analyzed with the flow cytometry assay. The expressions of the tumorrelated genes and proteins in A172 cells before and after cyclopamine treatment was detected with reverse transcriotionpolymerase chain reaction （RTPCR） and Western Blot. Cyclopamine affected A172 tumor growth in nude mice.ResultsCyclopamine inhibited the growth of A172 cells in a time and dosedependent manner. After cyclopamine treatment with different concentrations for 48 h, the apoptotic rates of A172 cells were increased in a dosedependent manner, which were significantly higher than those of the control group （P〈0.01）. Furthermore, cyclopamine downregulated the mRNA and protein levels of SMO and GLI1 in A172 cells. Cyclopamine inhibited A172 tumor growth in nude mice.ConclusionBlocking hedgehog signaling pathway inhibits human glioblastoma A172 cells growth in vivo and in vitro.
作者
薛立新
杨春福
XUE Lixin YANG Chunfu(Department of Neurosurgery, Zhijiang Hospital, Zhijiang 443200, Chin)
出处
《中华神经外科疾病研究杂志》
CAS
2017年第1期20-24,共5页
Chinese Journal of Neurosurgical Disease Research
