兰州大学机构库 >学院待认领
环王巴明对人前列腺癌Du145细胞的作用机制研究
Alternative TitleStudy on the mechanism of cyclopamine on human prostate cancer cell line Du145
孙全武
Thesis Advisor周逢海
2010-05-15
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword环王巴明 前列腺癌 Du145细胞 Hedgehog信号通路 细胞周期 凋亡
Abstract第一部分 环王巴明对Du145细胞增殖和细胞周期的影响 目的:研究Hedgehog信号通路阻断剂(环王巴明)对Du145细胞的抑制作用。 方法:不同浓度(1、10、50、100 μmol/L)环王巴明干预Du145细胞,分别在24、48、72 h后采用噻唑蓝比色法检测其对细胞的抑制作用;流式细胞术检测药物对细胞周期的影响;RT-PCR检测50 μmol/L环王巴明作用48 h后实验组和对照组细胞周期蛋白E(Cyclin E) mRNA表达水平差异。 结果:环王巴明对Du145细胞的抑制作用呈时效和量效依赖关系,当浓度﹥10μmol/L作用24 h后会显著抑制细胞增殖,10、50、100μmol/L浓度组对细胞的抑制率分别为:7.42%、12.70%、59.15%,与空白对照组相比,差异有统计学意义( P﹤0.05)。流式细胞术检测发现,当药物浓度达10 μmol/L以上,干预48 h后G1期细胞比例明显升高。10、50 μmol/L浓度组以及对照组的G1期百分比分别为:(60.13±2.75)%、(74.30±3.52)%、(52.17±2.21)%,差异有统计学意义( P﹤0.01),凋亡峰也逐渐增高;50 μmol/L环王巴明作用48 h后Du145细胞Cyclin E mRNA表达显著降低,与空白对照组相比降低约61.90% ( P﹤0.01)。 结论: 环王巴明可以抑制Du145细胞的增殖能力,其机制可能与下调Du145细胞Cyclin E mRNA表达水平,从而将Du145细胞阻滞于G1期。另外,环王巴明可以诱导Du145细胞凋亡。 关键词:环王巴明;前列腺癌;Du145细胞;Hedgehog信号通路;细胞周期 第二部分 环王巴明诱导DU145细胞凋亡的机制研究 目的: 研究低浓度胎牛血清培养下环王巴明诱导Du145细胞凋亡的作用,检测环王巴明对Du145细胞GLi-1和Bcl-2 mRNA表达水平的影响,探讨环王巴明诱导Du145细胞凋亡的作用机制。 方法:采用5、10μmol/L环王巴明处理Du145细胞,同时设立空白对照组。72h后透射电镜下观察细胞超微结构变化。流式细胞术检测药物作用后的细胞凋亡率变化,RT-PCR检测5、10μmol/L环王巴明作用72h后的实验组和对照组GLi-1、Bcl-2 mRNA表达水平差异; 结果:环王巴明作用72h后,在透射电镜下可见细胞核固缩,染色质边集,细胞膜微绒毛减少等典型的凋亡形态学改变。而对照组的细胞形态正常,细胞膜表面可见许多微绒毛。5μmol/L组的GLi-1、Bcl-2 mRNA表达减弱,但与对照组相比差异无统计学意义(P >0.05),10μmol/L环王巴明可以显著下调GLi-1、Bcl-2 mRNA表达,与空白及DMSO对照组相比差异有统计学意义(P﹤0.01); 结论:环王巴明可诱导人前列腺癌Du145细胞株凋亡,其作用机制可能与下调GLi-1和Bcl-2mRNA表达,从而活化细胞凋亡的线粒体途径有关。 关键词:环王巴明;前列腺癌;DU145细胞; 凋亡;
Other AbstractSection Ⅰ:The Effects of Cyclopamine on Cell Proliferation and Cell Cycle of Prostate Cancer Cell Line DU145 Abstract: 0bjective: To investigate the inhibitory effect of Hedgehog signal pathway blocker (cyclopamine) on Du145 cells. Methods: We interfered Du145 cells with cyclopamine at the concentration of 1, 10, 50 and 100 μmol/L and detected its inhibitory effect by MTT colorimetry assay at 24, 48 and 72 hours,as well as its effect on the cell cycle by flow cytometry, we also determined the difference in the mRNA expression of cyclin E between experiment and control group by RT-PCR at 48 hours after 50 μmol/L cyclopamine intervention. Results: Cyclopamine inhibited the Du145 cells in a time- and dose-dependent manner, with inhibition rates of 7.42%, 12.70% and 59.15% in the 10, 50 and 100 μmol/L groups respectively at 24 hours, significantly from that of the blank control group (P<0.05), It markedly suppressed the proliferation of the DU145 cells when cyclopamine over 10 μmol/L at 24 hours. Flow cytometry showed an obviously increased proportion of G1 phase cells when the concentration over 10 μmol/L after 48-hour intervention. with statistically significant differences from the proportion of G1 phase cells in the control, 10μmol/L and 50 μmol/L groups,which were (52.17±2.21)%, (60.13±2.75)% and (74.30±3.52)% respectively (P﹤0.01), and apoptotic peak was elevated with the increased concentration of cyclopamine; The cyclin E mRNA expression of Du145 cells was decreased by 61.90% at 48 hous after 50 μmol/L cyclopamine intervention as compared with the blank control group(P﹤0.01). Conclusion: Cyclopamine can inhibit the proliferation of Du145 cells, and the mechanism may be related with its effect of down-regulating the cyclin E mRNA expression of Du145 cells and blocking them in stage G1. Cyclopamine can also induce the apoptosis of Du145 cells. Key words: cyclopamine; prostate cancer; Du145 cell; Hedgehog signal pathway; cell cycle Section Ⅱ Study on the mechanism of cyclopamine induced apoptosis on human prostate cancer cell line Du145 Abstract: 0bjective: To investigate the effect of cyclopamine induced apoptosis on human prostate cancer cell line Du145, and its impact on GLi-1and Bcl-2 mRNA expression, make a preliminary explore to the mechanism of cyclopamine induced apoptosis on Du145; Methods: The morphological ultrastructural changes in Du145 cells after treatment with cyclopamine were studied using transmission electron m...
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223319
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
孙全武. 环王巴明对人前列腺癌Du145细胞的作用机制研究[D]. 兰州. 兰州大学,2010.
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