土贝母苷甲经miR-21/PDCD4途径诱导胶质瘤U251细胞凋亡  

Tubeimoside-1 Induces Apoptosis in Human Glioma Cell U251 Through miR-21/PDCD4 Pathway

作　　者：贾耿[1] 夏锡伟[1] 支枫[1] 彭亚[1] 邵耐远[1] 王强[1] 王穗暖[1] 杨伊林[1] 

机构地区：[1]苏州大学附属第三医院神经外科,江苏常州213003

出　　处：《现代生物医学进展》2016年第3期419-422,441,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(31071046;81302197);常州市社会发展项目(CS20092015;CS20102010);常州市卫生局重大课题(ZD200903;ZD201007);常州市科技局指导项目(CY20119004)

摘　　要：目的:研究土贝母苷甲(TBMS I)对胶质瘤U251细胞的抗肿瘤作用以及探究土贝母苷甲对miR-21及其靶基因PDCD4基因表达的影响。方法:U251细胞在体外进行培养,四甲基偶氮唑盐(MTT)法检测不同浓度的土贝母苷甲对细胞增殖的影响;Hoechst33258染色观察细胞核形态的变化;实时荧光定量PCR检测miR-21和PDCD4基因的表达情况;Western blot检测PDCD4蛋白的表达情况。结果:土贝母苷甲能够显著抑制U251细胞的增殖,其抑制作用呈剂量和时间依赖性。Hoechst33258染色观察到土贝母苷甲处理组细胞的细胞核形态表现出典型的凋亡特征。PCR结果显示:随着土贝母苷甲浓度增加,miR-21的表达逐渐降低(P<0.05),PDCD4基因表达显著增加(P<0.05)。Western blot结果提示:与阴性对照组相比,15、30μg/mL土贝母苷甲显著上调了PDCD4蛋白的表达(P<0.05)。结论:土贝母苷甲能够显著抑制人胶质瘤U251细胞的增殖并诱导细胞发生凋亡,其机制可能与下调miR-21的表达和上调PDCD4的表达有关。Objective： To study the antitumor effect of tubeimoside-1 on human glioma U251 cell line and explore the influence of tubeimoside-1 on the expression levels of miR-21 and programmed cell death 4（PDCD4）. Methods： U251 cells were cultured in vitro and treated with different concentrations of tubeimoside-1, cell viability was evaluated using MTT assay. Cell apoptosis was observed by Hoechst 33258 staining with a fluorescence microscope. The expression levels of miR-21 and PDCD4 m RNA were detected by real-time fluorescent quantitative PCR. Western blot analysis was used to detect the expression of PDCD4. Results： Tubeimoside-1 inhibited the proliferation of the U251 cells in a dose- and time-dependent manner. Cells exposed to tubeimoside-1 showed obviously changes of cellular ckaryomorphism which indicated apoptosis. Moreover, PCR analysis showed that after tubeimoside-1treatment, the expression of miR-21 tended to decrease gradually（P〈0.05）, down-regulation of miR-21 in U251 cell line resulted in increased expression of PDCD4（P〈0.05）. The expression of PDCD4 has also been enhanced by tubeimoside-1（15 and30 μg/mL） using Western blot analysis（P〈0.05）.Conclusions： Tubeimoside-1 significantly suppressed proliferation and induced apoptosis of U251 cells, the mechanism may be related to the down-regulation of miR-21 and up-regulation of PDCD4.

关 键 词：土贝母苷甲 胶质瘤 凋亡 MIR-21 PDCD4 

分 类 号：R-32[医药卫生]