β-葡萄糖苷酶激活苦杏仁苷诱导LoVo细胞凋亡及活性对Bax与bcl-2基因表达和Caspase-3的影响  

In vitro cytotoxity on colorectal carcinoma LoVo cells of a mygdalin following specific activation by β-glucosidase and influence on expr ession of bcl-2,bax gene and Caspase-3 activity

作　　者：连彦军[1] 陈道达[1] 黄韬[1] 柯茂林[1] 许天文[1] 郑勇斌[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院普外科,湖北武汉430022

出　　处：《肿瘤防治杂志》2005年第6期413-416,共4页China Journal of Cancer Prevention and Treatment

摘　　要：目的观察苦杏仁苷被β葡萄糖苷酶特异激活后对大肠癌LoVo细胞凋亡的影响。方法分别将不同浓度的苦杏仁苷加β葡萄糖苷酶作用于大肠癌LoVo细胞株24h,通过细胞形态学、DNA凝胶电泳、流式细胞仪观察细胞凋亡变化,RT PCR方法检测细胞bcl2、BaxmR NA表达变化,Caspase3试剂盒检测细胞Caspase3酶活性的变化。结果0.1～1.0mmol/L的苦杏仁苷与250nmol/L的β葡萄糖苷酶作用于细胞24h后,可见到以凋亡为主的形态变化细胞核固缩、染色质凝集及核碎裂、染色质片段化,琼脂糖凝胶电泳见DNA断裂成有规律的特征性梯状条带,流式细胞仪分析显示在G1期细胞前出现亚二倍体峰;RT PCR检测显示BaxmRNA表达升高,而bcl2mRNA表达无变化;Caspase3酶活性呈剂量依赖性增加。结论苦杏仁苷被β葡萄糖苷酶特异性激活后能导致LoVo细胞凋亡,这一过程中,Bax基因表达上调及Caspase3酶活性增加与LoVo细胞凋亡有关。OBJECTIVE:To study the effects on LoVo cells of amygdalin following specific activation by β-glucosidase.METHODS: Cytotoxity of amygdalin following specific activity by β-glucosidas e on LoVo cells were investigated by morphology, agarose gel electrophoresis and flow cytometry, the levels of mRNA of genes bcl-2 and bax were determined by r everse transcription-polymerase chain reaction respectively, and caspase-3 act ivity was measured by fluorescent assay kits.RESULTS:After 24 hours exposure to 0.1-1.0 mmol/L amygdalin and 250 nmol/L β-glucosidase, LoVo cells presented some typical feasures of apoptosis: nucler shrinkage, chro matin condensation and DNA fragmentation. agarose gel electrophoresis appeared m arked DNA ladder, flow cytometry analysis showed a typical subdiploid peak befor e G_0/G_1 phase, the levels of bax mRNA was up-regulated, while the level of bcl-2 mRNA was not affected, caspase-3 activity increased in a dose-dependent mode.CONLUSIONS:Amygdalin, once specificly activited by β-glucosidase, can induce LoVo cells apoptosis, the mechanism of inducing apop tosis is related to up-regulation of bax gene expression and caspase-3 activit y.[

关 键 词：苦杏仁苷 Β-葡萄糖苷酶 大肠癌 凋亡 CASPASE-3 

分 类 号：R735.34[医药卫生—肿瘤]