姜黄素MCF-7细胞增殖不同浓度姜黄素作用MCF-7MTT测细胞.PPTVIP

Proteomic identification of differentially expressed proteins in curcumin-treatedMCF-7 cells 作者：H.Y. Fang 刊名：Phytomedicine18 (2011) 697–703 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 IC50value of CM in MCF-7 cells was determined as 47.42μM. 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 47.42μM 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 Compar