香加皮单体成分宝藿苷I对食管癌细胞增殖及凋亡的影响  

Effect of baohuoside Ⅰ from Cortex Periplocae on proliferation and apoptosis of esophageal carcinoma cells

作　　者：王丽芳[1,2] 单保恩[1] 刘丽华[1] 任凤芝[3] 单铁强[1] 

机构地区：[1]河北医科大学第四医院科研中心,石家庄050011 [2]河北医科大学第二医院检验科,石家庄050000 [3]华北制药集团新药研究开发有限责任公司,石家庄050015

出　　处：《肿瘤》2009年第2期123-126,共4页Tumor

基　　金：国家自然科学基金资助项目(编号:30772752);河北省自然科学基金资助项目(编号:C2008000952)

摘　　要：目的:研究香加皮单体成分宝藿苷Ⅰ在体内外对人食管癌细胞Eca109的增殖抑制作用及其对细胞凋亡的影响。方法:采用柱层析和高效液相色谱等逐步分离法对香加皮抗肿瘤活性成分进行分离和纯化,应用薄层层析和电喷质谱分析进行成分鉴定;采用MTT法分析不同浓度宝藿苷Ⅰ对食管癌细胞株Eca109增殖的抑制作用;应用FCM分析细胞周期变化和凋亡率变化;皮下注射Eca109细胞建立裸鼠移植瘤动物模型,通过肌肉注射给药检测宝藿苷Ⅰ的体内抗肿瘤效果;Western印迹法检测移植瘤组织凋亡相关基因survivin的表达变化。结果:宝藿苷Ⅰ可明显抑制Eca109细胞的增殖(P<0.05),并呈浓度及时间依赖性。经宝藿苷Ⅰ作用48h后,随着药物浓度的增加(12.5、25.0和50.0μg/mL),Eca109细胞G0/G1期明显增加(P<0.05),S期和G2/M期细胞明显减少(P<0.05);经宝藿苷Ⅰ作用后,Eca109细胞的凋亡率随药物作用时间的延长(24、48和72h)和宝藿苷浓度的增加(0、12.5、25.0和50.0μg/mL)而明显升高(P<0.01);Eca109裸鼠移植瘤经宝藿苷Ⅰ(15mg/kg)治疗后,肿瘤生长受到明显抑制(P<0.01),生长抑制率达(60.9±0.16)%;survivin的蛋白表达明显降低(P<0.05)。结论:香加皮单体成分宝藿苷Ⅰ不仅在体外对Eca109细胞的增殖具有显著的抑制作用,在体内也有较好的抗食管癌效果。其抗瘤机制可能与阻滞Eca109细胞周期发展和诱导凋亡相关基因survivin的表达降低有关。Objective ： To investigate the inhibitory effect of baohuoside Ⅰ from Cortex Periplocae on the proliferation and apoptosis of esophageal carcinoma in vivo and in vitro. Methods ： Baohuoside Ⅰ was purified preliminarily using stepwise analysis method. Ⅰt was isolated and purified further by column chromatography and high pressure liquid chromatography. The components were identified by thin layer chromatogram and electrospray ionization-mass spectrometry （ESI-MS）. The inhibitory effect of baohuoside Ⅰ on proliferation of Eca109 tumor cell line was examined by using MTT assay; the changes of cell cycle and the apoptotic ratio were analyzed with flow cytometry. Eca109 ceils were injected subcutaneously into nude mice to establish xenografted tumor model. Baohuoside Ⅰ was intramuscularly injected into nude mice to observe its anti-tumor effects in vivo. The expression of tissue apoptosis-related gene survivin in xenografted tumor tissues was measured by Western blotting. Results： Baohuoside Ⅰ significantly inhibited the proliferation of Eca109 tumor cells in a concentration- and time-dependent manner （P 〈 0.05）. After treatment with baohuoside Ⅰ at 12.5, 25.0, 50.0μg/ mL for 48 h, the cell proportion in G0/G1 phase was gradually increased （P 〈0.05） and the cells in S and GJM phase were significantly decreased （P 〈0.05）. The apoptotic rate of Eca109 cells was significantly increased by Baohuoside Ⅰ in a time-dependent （24, 48, and 72 h） and concentration-dependent （0, 12.5, 25.0, and 50.0 μg/mL） manner. Baohuoside Ⅰ at 15 mg/kg significantly inhibited the growth of xenografted tumor in nude mice （P 〈 0.01 ） with the inhibitory ratio of （60.9 ± 0. 16 ）%. The expression of survivin protein was significantly decreased （P 〈 0.05 ）. Conclusion： The component of Cortex Periplocae, Baohuoside Ⅰ , had significant inhibitory effects on the proliferation of Eca109 cells in vitro and has better anti-esophageal carcinoma effects in vivo. The mechanism ma

关 键 词：食管肿瘤 模型  动物 细胞  ECA109 香加皮 宝藿苷Ⅰ 

分 类 号：R735.1[医药卫生—肿瘤] R73-36[医药卫生—临床医学]