脱氢枞胺对氟苯甲醛对人肝癌细胞的抑制作用  

Inhibitory Effect of Dehydroabietylamine-Fluorobenzaldehyde on Human Hepatocellular Carcinoma Cells

作　　者：刘玲[1] 李富[2] 何玲[2] 饶小平[3] 宋湛谦 

机构地区：[1]河南科技大学医学院药理教研室,洛阳471003 [2]中国药科大学药理教研室,南京210009 [3]中国林业科学院林产化学工业研究所,南京210042

出　　处：《中国药学杂志》2013年第4期269-274,共6页Chinese Pharmaceutical Journal

摘　　要：目的观察脱氢枞胺对氟苯甲醛[1-(7-异丙基-1,4a-二甲基-1,2,3,4,4a,9,10,10a-八氢菲基-1-yl)-N-(4-氟苯亚甲基)甲胺]对人肝癌细胞的抑制作用及其分子机制。方法运用四甲基偶氮唑蓝、倒置显微镜、流式细胞仪、Western blot、荧光分光光度法等分别检测脱氢枞胺对氟苯甲醛对肝癌细胞增殖的抑制作用、观察凋亡细胞细胞核的形态变化、检测细胞凋亡和线粒体膜电位变化、凋亡相关蛋白Bcl-2、Bax、p53、cyt-c的表达和caspase-3活性的影响。建立H22小鼠体内移植实体瘤模型,观察脱氢枞胺对氟苯甲醛对瘤体质量的影响。结果四甲基偶氮唑蓝法显示脱氢枞胺对氟苯甲醛能显著抑制肝癌细胞株(SMMC-7721、HepG2和BEL-7402细胞)的增殖,IC50值分别为SMMC-7721(44.47±2.15)μmol·L-1、HepG2(52.83±2.25)μmol·L-1和BEL-7402(48.64±1.76)μmol·L-1。倒置显微镜下发生细胞皱缩变圆等变化,细胞凋亡率明显增加,线粒体膜电位降低,Bcl-2表达减少,Bax、p53和Cyt c表达增加,caspase-3活力增加,且均具有浓度依赖性。小鼠H22肝癌模型中,脱氢枞胺对氟苯甲醛对肿瘤的生长也有一定的抑制作用。结论体内外实验显示脱氢枞胺对氟苯甲醛能明显抑制肿瘤细胞的生长,诱导肿瘤细胞凋亡,可能是通过凋亡线粒体信号通路而实现的。OBJECTIVE To examine the anti-proliferation effects of a novel derivative of dehydroabietylamine, dehydroabietyl-amine-fluorobenzaldehyde [ 1-（ 7-isopropyl-1,4a-dimethyl-1,2,3,4,4a, 9, 10, 10a-octahydrophenanthren-l-yl ） -N-（ 4-fluorobenzyli- dene）methanamine,DHAA-F] , on human hepatoeellular carcinoma cells and to expolore its molecular mechanism. METHODS MTI assay was adopted to detect the proliferation status of the cells treated with DHAA-F ; cellular apoptosis and reduction of mitoehondria membrane potential （ AΨm ） were analyzed using flow cytometry （FCM） ; Western blotting assay was used to evaluate the release of Cyt c, and the expressions of p53, Bcl-2 and Bax protein; and the caspase-3 activity was determined with fluorescence spectrophotometry. To evaluate the anti-tumor effect of DHAA-F in vivo, mouse model beating inoculated H22 tumor was established. RESULTS DHAA-F strongly inhibited human hepatoma cells proliferation. The IC50 value of DHAA-F was （44.47 -＋ 2. 15 ） μmol·L- 1 for SMMC- 7721, （48.64 ＋ 1.76） μmol·L-1 for Bel-7402, and （52. 83 ＋2. 25） μmol·L-l for HepG2. DHAA-F displayed a significant inhibieffect on the growth of SMMC-7721 cells in a dose- and time-dependent manner. When SMMC-7721 cells were pretreated with DHAA-F for 24 h, the apoptosis rate significantly increase and the mitochondrial membrane potential significantly decreased. Western blotting assay showed significant decrease of Bcl-2 protein expression and increase of Bax, p53 protein expression, cytosol Cyt e level and caspase-3 activity. DHAA-F could significantly reduce tumor weight in the H22 solid tumor mouse model in vivo. CONCLUSION These findings suggest that DHAA-F has potent antitumor activity both in vivo and in vitro and the mechanism may be related to the apoptosis induced by DHAA-F through a mitoehondrial pathway.

关 键 词：1-(7-异丙基-1 4a-二甲基-1 2 3 4 4a 9 10 10a-八氢菲基-1-yl)-N-(4-氟苯亚甲基)甲胺 脱氢枞胺对氟苯甲醛 凋亡 线粒体 肝癌 

分 类 号：R965[医药卫生—药理学]