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

1、Proteomic identification of differentially expressed proteins in curcumin-treatedMCF-7 cells石文贵作者：作者：H.Y. Fang刊名：刊名：Phytomedicine18 (2011) 697703单位：单位：Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College目录目录1 1研究背景研究背景2 2实验方法实验方法3 3研究结果研究结果4

2、4结果讨论结果讨论一一. .研究背景研究背景1.1.姜黄素及其生物功效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。一一. .研究背景研究背景2.2.蛋白质组学研究的优势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.实验方法姜黄素姜黄素MCF-7细胞增细胞增殖实验殖实验不同浓度姜黄不同浓度姜黄素作用素作用

3、MCF-7MTT测测细胞增细胞增殖殖最佳作最佳作用浓度用浓度细胞凋细胞凋亡实验亡实验凋亡细胞形态凋亡细胞形态变化变化凋亡细胞数量凋亡细胞数量的变化的变化凋亡细胞所处凋亡细胞所处的细胞周期的细胞周期Hoechst 荧荧光染色法光染色法Annexin V/PI 双染色法双染色法流式细胞仪流式细胞仪双向电双向电泳泳蛋白质蛋白质组学组学MALDI-TOF-MS根据蛋白种根据蛋白种类设计引物类设计引物变化的变化的蛋白蛋白Real-timePCR基因基因表达表达三三. .实验结果实验结果1.细胞增殖实验结果Antiproliferative effects of curcumin and cisplati

4、n in MCF-7 cells. Cell via-bility was measured with the MTT colorimetric assay after treatment for 48h at indicated concentrations. The IC50value of CM in MCF-7 cells was determined as 47.42M.三三. .实验结果实验结果2.细胞凋亡实验结果Cells were stained by Hoechst 33258 (a) Control and (b), treatment with CM at 47.42 M

5、 for 48h (left). The bar graph shows the average number of Hoechst staining cells for control and CM (right) from three microscopyviews (*p0.01).三三. .实验结果实验结果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

6、cluster is of necrotic cells.三三. .实验结果实验结果2.细胞凋亡实验结果The proportion of MCF-7 cells in S phase at 47.42M 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 cyclepro

7、gression三三. .实验结果实验结果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三三. .实验结果实验结果4. MALDI-TOF-MSControlCM三三. .实验结果实验结果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TD