Proteomic identification of differentially expressed proteins in curcumin-treated文獻講解

1、Proteomic identification of differentially expressed proteins in curcumin-treatedMCF-7 cells石文貴作者：作者：H.Y. Fang刊名：刊名：Phytomedicine18 (2011) 697703單位：單位：Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College目錄目錄1 1研究背景研究背景2 2實驗方法實驗方法3 3研究結(jié)果研究結(jié)果4

2、4結(jié)果討論結(jié)果討論一一. .研究背景研究背景1.1.姜黃素及其生物功效姜黃素及其生物功效 姜黃素是從姜黃中提取的一種多酚類化合 物，具有抗炎、抗氧化、抗癌等多種功效。其與紫杉醇結(jié)合對乳腺癌具有很好的治療作用，但姜黃素抗癌的生物學(xué)機理還不明確。一一. .研究背景研究背景2.2.蛋白質(zhì)組學(xué)研究的優(yōu)勢蛋白質(zhì)組學(xué)研究的優(yōu)勢 藥物刺激細胞后，蛋白的含量和性質(zhì)會發(fā)生變化，而蛋白質(zhì)組學(xué)技術(shù)能對這些蛋白很好地進行定性定量，本文就是利用蛋白質(zhì)組學(xué)技術(shù)，對姜黃素作用后的MCF-7細胞后，蛋白質(zhì)含量和性質(zhì)的變化經(jīng)行研究。 二.實驗方法姜黃素姜黃素MCF-7細胞增細胞增殖實驗殖實驗不同濃度姜黃不同濃度姜黃素作用素作用

3、MCF-7MTT測測細胞增細胞增殖殖最佳作最佳作用濃度用濃度細胞凋細胞凋亡實驗亡實驗凋亡細胞形態(tài)凋亡細胞形態(tài)變化變化凋亡細胞數(shù)量凋亡細胞數(shù)量的變化的變化凋亡細胞所處凋亡細胞所處的細胞周期的細胞周期Hoechst 熒熒光染色法光染色法Annexin V/PI 雙染色法雙染色法流式細胞儀流式細胞儀雙向電雙向電泳泳蛋白質(zhì)蛋白質(zhì)組學(xué)組學(xué)MALDI-TOF-MS根據(jù)蛋白種根據(jù)蛋白種類設(shè)計引物類設(shè)計引物變化的變化的蛋白蛋白Real-timePCR基因基因表達表達三三. .實驗結(jié)果實驗結(jié)果1.細胞增殖實驗結(jié)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. .實驗結(jié)果實驗結(jié)果2.細胞凋亡實驗結(jié)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. .實驗結(jié)果實驗結(jié)果2.細胞凋亡實驗結(jié)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. .實驗結(jié)果實驗結(jié)果2.細胞凋亡實驗結(jié)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. .實驗結(jié)果實驗結(jié)果3.雙向電泳結(jié)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. .實驗結(jié)果實驗結(jié)果4. MALDI-TOF-MSControlCM三三. .實驗結(jié)果實驗結(jié)果3.Real-time PCR結(jié)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.討論1.姜黃素抑制乳腺癌細胞的增殖，一個主要原因是引起了細胞的凋亡。2.通過質(zhì)譜分離的7種上調(diào)蛋白和5中下調(diào)蛋白，對這12種蛋白的研究將是解釋姜黃素治療乳腺癌的關(guān)鍵3.比如TD