贾耿 夏锡伟 支枫 彭亚 邵耐远 王强 王穗暖 杨伊林.土贝母苷甲经miR-21/ PDCD4 途径诱导胶质瘤U251细胞凋亡[J].,2016,16(3):419-422 |
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 ThroughmiR-21/PDCD4 Pathway |
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DOI: |
中文关键词: 土贝母苷甲 胶质瘤 凋亡 miR-21 PDCD4 |
英文关键词: Tubeimoside-1 Glioma Apoptosis miR-21 PDCD4 |
基金项目: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) |
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中文摘要: |
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、30 滋g/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。 |
英文摘要: |
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 death4 (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 mRNA 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 ug/mL) using Western blot analysis(P<0.05)Conclusion:Tubeimoside-1 significantly suppressed proliferation and induced apoptosis of U251 cells, the mechanismmay be related to
the down-regulation of miR-21 and up-regulation of PDCD4. |
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