Proteomic-identification-of-differentially-expressed-proteins-in-curcumin-treated文献讲解

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,Proteomic identification of differentially expressed proteins in curcumin-treatedMCF-7 cells,石文贵,作者：,H.Y.Fang,刊名：,Phytomedicine18(2011)697703,单位：,Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College,目录,1,研究背景,2,实验方法,3,研究结果,4,结果讨论,一,.,研究背景,1.,姜黄素及其生物功效,姜黄素是从姜黄中提取的一种多酚类化合 物，具有抗炎、抗氧化、抗癌等多种功效。其与紫杉醇结合对乳腺癌具有很好的治疗作用，但姜黄素抗癌的生物学机理还不明确。,一,.,研究背景,2.,蛋白质组学研究的优势,药物刺激细胞后，蛋白的含量和性质会发生变化，而蛋白质组学技术能对这些蛋白很好地进行定性定量，本文就是利用蛋白质组学技术，对姜黄素作用后的,MCF-7,细胞后，蛋白质含量和性质的变化经行研究。,二,.,实验方法,姜黄素,MCF-7,细胞增殖实验,不同浓度姜黄素作用,MCF-7,MTT,测细胞增殖,最佳作用浓度,细胞凋亡实验,凋亡细胞形态变化,凋亡细胞数量的变化,凋亡细胞所处的细胞周期,Hoechst,荧光染色法,Annexin,V/PI,双染色法,流式细胞仪,双向电泳,蛋白质组学,MALDI-TOF-MS,根据蛋白种类设计引物,变化的蛋白,Real-,timePCR,基因表达,三,.,实验结果,1.,细胞增殖实验结果,Antiproliferative,effects of curcumin and,cisplatin,in MCF-7 cells.Cell via-,bility,was measured with the MTT colorimetric assay after treatment for 48h at indicated concentrations.The IC,50,value of CM in MCF-7 cells was determined as 47.42,M.,三,.,实验结果,2.,细胞凋亡实验结果,Cells were stained by Hoechst 33258(a)Control and(b),treatment with CM at 47.42M for 48h(left).The bar graph shows the average number of Hoechst staining cells for control and CM(right)from three microscopy,views(*p0.01).,三,.,实验结果,2.,细胞凋亡实验结果,The cell cluster in the lower left area represents live,cells.The,cells in the right areas are apoptotic cells.The upper-left cell cluster is of necrotic cells.,三,.,实验结果,2.,细胞凋亡实验结果,The proportion of MCF-7 cells in S phase at 47.42,M of CM,，,increased from 43.6%to 54.7%after 48h exposure with a con-,comitant,increase of G2/M cells from 4.93%to 9.47%.The result,，,Indicated that CM treat,ment,hampered S andG2/M,phasescell,cycleprogression,三,.,实验结果,3.,双向电泳结果,A total of 53 protein spots were found to be differentially expressed between curcumin-treated groups and control groups,of which 14 spots were down-regulated and 39 spots up-regulated.Twelve proteins were successfully identified by MALDI-TOF-MS,三,.,实验结果,4.,MALDI-TOF-MS,Control,CM,三,.,实验结果,3.Real-time PCR,结果,Comparative expression of genes as determined by RT-PCR.,The ratio of the control group was regard as one.*p0.05 compared to the control,四,.,讨论,1.,姜黄素抑制乳腺癌细胞的增殖，一个主要原因是引起了细胞的凋亡。,2.,通过质谱分离的,7,种上调蛋白和,5,中下调蛋白，对这,12,种蛋白的研究将是解释姜黄素治疗乳腺癌的关键,3.,比如,TDP-43,使,pRb,磷酸化失活，姜黄素使,TDP-43,表达下降，激活了,pRb,途径，引起了细胞凋亡。,4.ERP29,是一种内质网结合蛋白。它能够激活与细胞凋亡有关的蛋白。,ERP29,的过表达使得细胞停留在,G,0,/G,1,，抑制细胞的增殖。,谢谢大家！,