芹菜素对人胶质瘤SHG-44细胞株增殖抑制和诱导凋亡的实验研究

发布时间：2018-05-28 05:25

本文选题：芹菜素 + 胶质瘤　；参考：《江苏大学》2017年硕士论文

【摘要】：目的探讨不同浓度芹菜素对胶质瘤SHG-44细胞增殖抑制和诱导凋亡的作用。方法我们分别以不同浓度(20、40、80μmol/L)的芹菜素处理SHG-44胶质瘤细胞24、48、72 h,采用MTT法检测不同浓度芹菜素对胶质瘤细胞增殖抑制率的影响。运用PI单染流式细胞仪检测法观察芹菜素处理对胶质瘤24 h后,细胞周期的分布情况。采用Hoechst 33258、Tunel染色法、AnnexinV-FITC/PI双染流式细胞仪检测技术探究不同浓度芹菜素对胶质瘤细胞凋亡的影响。通过蛋白印迹方法(Western blot)观察细胞周期相关蛋白CDK6、CDC25A及凋亡途径相关蛋白Bax、Bcl-2的表达量的变化。结果芹菜素对SHG-44细胞的增殖抑制作用显著,且随药物作用时间和作用浓度的增加而增强。细胞周期检测显示芹菜素使胶质瘤细胞发生G2/M期阻滞。Hoechst33258免疫荧光染色显示芹菜素处理组胶质瘤细胞可见染色质固缩、核断裂和调亡小体等典型细胞凋亡表现,TUNEL免疫荧光染色显示凋亡细胞出现绿色荧光。流式细胞仪分析结果显示,随着芹菜素处理浓度的增加,SHG-44细胞的凋亡率逐渐增高,不同浓度芹菜素作用24 h的结果显示该药物能呈浓度依赖性地诱导SHG-44细胞凋亡。芹菜素处理组细胞的CDK6、CDC25A蛋白表达量逐渐下降,Bax蛋白表达量逐渐增加,Bcl-2蛋白表达量逐渐降低,Bax/Bcl-2比值逐渐增大。结论在体外实验中,芹菜素可抑制胶质瘤SHG-44细胞增殖,并呈时间和剂量依赖性。同时芹菜素可诱导胶质瘤SHG-44发生凋亡;芹菜素可使胶质瘤细胞发生G2/M期阻滞,其机制可能与CDK6、CDC25A表达的变化有关;芹菜素可通过Bcl-2相关途径诱导胶质瘤SHG-44细胞凋亡。
[Abstract]:Objective to investigate the effects of apigenin at different concentrations on the proliferation inhibition and apoptosis induction of glioma SHG-44 cells. Methods SHG-44 glioma cells were treated with apigenin at different concentrations of 4080 渭 mol / L for 2448 ~ 72 h. The effects of apigenin at different concentrations on the proliferation inhibition rate of glioma cells were detected by MTT assay. The distribution of cell cycle in gliomas treated with apigenin for 24 h was observed by Pi single staining flow cytometry. The effect of apigenin at different concentrations on apoptosis of glioma cells was investigated by Hoechst 33258 Tunel staining and Annexin V-FITC / Pi double staining flow cytometry. The expression of cell cycle associated protein CDK6, CDC25A, and apoptosis-related protein BaxanBcl-2 were observed by Western blot. Results apigenin significantly inhibited the proliferation of SHG-44 cells and increased with the time and concentration of action. Cell cycle analysis showed that apigenin caused G2 / M phase arrest. Hoechst33258 immunofluorescence staining showed chromatin pyknosis in glioma cells treated with apigenin. Tunel immunofluorescence staining showed green fluorescence in apoptotic cells such as nuclear fragmentation and apoptotic bodies. Flow cytometry analysis showed that the apoptosis rate of SHG-44 cells increased with the increase of apigenin concentration. The results of 24 h treatment with different concentrations of apigenin showed that the drug could induce apoptosis of SHG-44 cells in a concentration-dependent manner. In apigenin treated group, the expression of CDK6 and CDC25A protein decreased gradually, the expression of Bax protein increased gradually, the expression of Bcl 2 protein decreased gradually and the ratio of Bax / Bax protein gradually increased. Conclusion apigenin can inhibit the proliferation of glioma SHG-44 cells in a time and dose-dependent manner. Apigenin can induce apoptosis of glioma SHG-44, apigenin can block G2 / M phase of glioma cells, and its mechanism may be related to the change of CDK6, CDC25A expression. Apigenin can induce apoptosis of glioma SHG-44 cells through Bcl-2 related pathway.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.41

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