姜黄素对人前列腺癌DU-145细胞TGF-β/Smads信号传导通路的影响
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摘要
目的：探讨中药提取物姜黄素对人雄激素非依赖性前列腺癌DU-145细胞TGF-β/Smads信号传导通路的影响,从而探讨姜黄素抑制DU-145细胞的机理。
方法：采用细胞培养技术,将姜黄素配制成12.5μmol/l、6.25μmol/l以及3.125μmol/l三个不同浓度组的溶液,分别处理DU-145细胞48小时。以光镜观察DU-145细胞形态变化,以MTT法检测DU-145细胞生长情况,以ELLISA法和RT-PCR法检测TGF-β,SMAD2, SMAD4和SMAD7的浓度和表达情况。
结果：姜黄素溶液作用DU-145细胞48小时后,出现了细胞凋亡的形态学改变,表现为数量减少、体积缩小、染色质凝集、核固缩；DU-145细胞生长受到抑制,呈一定的量效关系；ELLISA法检测DU-145细胞TGF-β,SMAD2和SMAD4的吸光度OD值(代表上述物质浓度)均不同程度增加,而SMAD7的OD值不同程度减少；RT-PCR法检测DU-145细胞TGF-β,SMAD2和SMAD4基因表达均不同程度增加,而SMAD7的基因表达不同程度减少。两种检测法得出结果一致。
结论：姜黄素通过对TGF-β及下游蛋白Smad2、Smad4、Smad7表达的影响,进而激活或调节TGF-β/Smads信号传导通路,这可能是姜黄素促进DU-145细胞凋亡和抑制其生长的重要分子机制。
Objectives:To observe the efficacy Curcumine having on TGF-(3/Smads pathway of human DU-145 prostatic cancer cells in hormone-independent prostatic cancer, and to accordingly observe the principle of it restraining DU-145 cells.
Methods:Cell cultivation was adopted with different groups using 12.5μmol/l,6.25μmol/l and 3.125μmol/l Curcumine concentrations on DU-145 cells for 48 hours each. The DU-145 cells'morphology was observed under optic microscope. The multiplication of DU-145 cells was measured by MTT method. Both ELLISA and RT-PCR methods was used to measure the concentrations and expressions of TGF-β,SMAD2, SMAD4 and SMAD7.
Results:DU-145 cells'morphologic changes were shown with number decreasing, volume reduction, chromatin coagulation and nucleus contraction after 48 hours'influence Curcumine concentrations had on them. A certain quantity-effect relation was shown in each concentration group. The values of OD of TGF-β,SMAD2 and SMAD4 were increased by the concentration groups measured with ELLISA but SMAD7 were lowed. The expressions of TGF-β,SMAD2 and SMAD4 were increased by the concentration groups measured with RT-PCR but SMAD7 were lowed. The two methods of measurement showed the same results.
Conclusions:To intervention TGF-β, SMAD2, SMAD4 and SMAD7 of human DU-145 prostatic cancer cells,curcumine can activation or regulate TGF-β/Smads pathway of them,what could be the important mechanism of curcumine restraining DU-145 cells
方法：采用细胞培养技术,将姜黄素配制成12.5μmol/l、6.25μmol/l以及3.125μmol/l三个不同浓度组的溶液,分别处理DU-145细胞48小时。以光镜观察DU-145细胞形态变化,以MTT法检测DU-145细胞生长情况,以ELLISA法和RT-PCR法检测TGF-β,SMAD2, SMAD4和SMAD7的浓度和表达情况。
结果：姜黄素溶液作用DU-145细胞48小时后,出现了细胞凋亡的形态学改变,表现为数量减少、体积缩小、染色质凝集、核固缩；DU-145细胞生长受到抑制,呈一定的量效关系；ELLISA法检测DU-145细胞TGF-β,SMAD2和SMAD4的吸光度OD值(代表上述物质浓度)均不同程度增加,而SMAD7的OD值不同程度减少；RT-PCR法检测DU-145细胞TGF-β,SMAD2和SMAD4基因表达均不同程度增加,而SMAD7的基因表达不同程度减少。两种检测法得出结果一致。
结论：姜黄素通过对TGF-β及下游蛋白Smad2、Smad4、Smad7表达的影响,进而激活或调节TGF-β/Smads信号传导通路,这可能是姜黄素促进DU-145细胞凋亡和抑制其生长的重要分子机制。
Objectives:To observe the efficacy Curcumine having on TGF-(3/Smads pathway of human DU-145 prostatic cancer cells in hormone-independent prostatic cancer, and to accordingly observe the principle of it restraining DU-145 cells.
Methods:Cell cultivation was adopted with different groups using 12.5μmol/l,6.25μmol/l and 3.125μmol/l Curcumine concentrations on DU-145 cells for 48 hours each. The DU-145 cells'morphology was observed under optic microscope. The multiplication of DU-145 cells was measured by MTT method. Both ELLISA and RT-PCR methods was used to measure the concentrations and expressions of TGF-β,SMAD2, SMAD4 and SMAD7.
Results:DU-145 cells'morphologic changes were shown with number decreasing, volume reduction, chromatin coagulation and nucleus contraction after 48 hours'influence Curcumine concentrations had on them. A certain quantity-effect relation was shown in each concentration group. The values of OD of TGF-β,SMAD2 and SMAD4 were increased by the concentration groups measured with ELLISA but SMAD7 were lowed. The expressions of TGF-β,SMAD2 and SMAD4 were increased by the concentration groups measured with RT-PCR but SMAD7 were lowed. The two methods of measurement showed the same results.
Conclusions:To intervention TGF-β, SMAD2, SMAD4 and SMAD7 of human DU-145 prostatic cancer cells,curcumine can activation or regulate TGF-β/Smads pathway of them,what could be the important mechanism of curcumine restraining DU-145 cells
引文
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