乌贼墨多肽诱导人前列腺癌DU-145细胞凋亡的机制研究  

Mechanism of Sepia Ink Polypeptide-induced Apoptosis in DU-145 Prostate Cancer Cells

作　　者：景奕文[1] 杨最素[1] 黄芳芳[1] 郁迪[1] 丁国芳[1] 

机构地区：[1]浙江海洋学院食品与医药学院、浙江省海洋生物医用制品工程技术研究中心,浙江舟山316004

出　　处：《现代食品科技》2014年第9期1-6,共6页Modern Food Science and Technology

基　　金：国家自然科学基金面上项目(81273429);浙江省科技厅重大专项(2010C13009;2011C02003);浙江省自然科学基金立项(LY12C20005;LY12C20008);舟山市科技计划项目(2012C23023)

摘　　要：本文探讨了乌贼墨多肽(SHP)诱导DU-145细胞凋亡机制。采用CCK-8法检测SHP对DU-145细胞增殖的影响;采用HE染色和AO/EB荧光染色观察DU-145细胞的形态学的变化;采用流式细胞术检测细胞早期凋亡率;并通过Western Blotting检测细胞中p53、Bcl-2、Bax、Caspase-3、VEGF的蛋白表达变化。结果表明,SHP对DU-145细胞的增殖具有明显的抑制作用且呈现剂量和时间依赖性;SHP作用后的DU-145细胞出现凋亡的形态学特征;流式细胞术结果显示,随着SHP浓度和作用时间的增加,DU-145细胞的早期凋亡率从12.25%增加到34.20%;Western Blotting结果显示,当SHP作用24 h后,VEGF、Bcl-2蛋白表达量降低,p53、Bax、Caspase-3蛋白表达量增加。综上可知,SHP能够诱导DU-145细胞凋亡,其机制有可能是通过激活抑癌基因p53,下调Bcl-2/Bax比例;下调VEGF,激活凋亡蛋白酶Caspase-3,诱发凋亡级联反应来实现的。In this study, the mechanism of apoptosis induced by sepia ink polypeptide （SHP） in DU-145 prostate cancer cells was explored. The effect of SHP on the proliferation of DU-145 cells was examined by the Cell Counting Kit-8 （CCK-8） assay. Typical morphological changes in DU-145 cells were observed with hematoxylin and eosin （HE） and acridine orange/ethidium bromide （AO/EB） staining. The early-stage apoptosis rate was measure using flow cytometry （FCM）, and the changes in the expression of apoptosis-related genes （p53, Bcl-2, Bax, Caspase-3, and VEGF） were evaluated via western blotting. The results showed that SHP significantly inhibited the proliferation of DU-145 cells in a time-and dose-dependent manner. DU-145 cells developed morphological features of apoptosis after treated with SHP.FCM studies revealed that the early-stage apoptosis rate of DU-145 cells increased from 12.25% to 34.20% with increasing SlIP concentration and duration of treatment. Western blotting results showed that after 24 h treatment with SHP, the expression of anti-apoptotic proteins Bcl-2 and VEGF decreased, while the expression of p53, Bax, and Caspase-3 increased. Collectively, these results suggest that SHY induced apoptosis in DU-145 cells. The mechanism might involve the decrease in Bcl-2/Bax expression ratio by activation of the tumor suppressor gene p53. Moreover, VEGF expression was down regulated, and apoptotic protease Caspase-3 was activated, thus triggering the apoptosis cascade reaction.

关 键 词：乌贼墨 多肽 凋亡 

分 类 号：R737.25[医药卫生—肿瘤]