二烯丙基二硫诱导人鼻咽癌CNE2细胞凋亡及其可能的分子机制  

Apoptosis of nasopharyngeal cancer cells line CNE2 induced by diallyl disulfide and its possible molecular mechanism

作　　者：唐海林[1] 宋颖[1] 唐荣军[1] 周秀田[1] 曾希[1] 苏琦[1] 

机构地区：[1]南华大学肿瘤研究所,湖南衡阳421001

出　　处：《中国癌症杂志》2007年第8期598-602,共5页China Oncology

基　　金：湖南省教育厅科学研究项目基金资助项目(No:06C695)

摘　　要：背景与目的:二烯丙基二硫(DADS)是大蒜中的一种脂溶性单体化合物,对多种肿瘤细胞有促凋亡作用。本实验旨在探讨DADS抑制人鼻咽癌CNE2细胞增殖与诱导凋亡的生物学效应及可能的分子机制。方法:分别采用MTT、光学显微镜、流式细胞仪、DNA琼脂糖凝胶电泳与Western blot检测DADS抑制CNE2细胞增殖与诱导凋亡作用及Bcl-2、Bax和半胱氨酸天冬氨酸蛋白酶(Caspase3)蛋白的表达。结果:MTT结果显示,50、100、200及400μmol/LDADS作用CNE2细胞48h,抑制率分别为3.66%、14.25%、29.66%及61.28%,呈浓度依赖性(P<0.05)。形态学观察发现,DADS处理24h后,部分CNE2细胞变圆,体积明显缩小,胞质强嗜酸性,核固缩、深染,核染色质积聚至核膜周边,呈现凋亡细胞形态学改变。流式细胞术分析显示,50、100、200和400μmol/LDADS作用24h,凋亡率分别为(10.8±1.3)%、(14.8±1.1)%、(21.7±2.0)%及(29.8±2.6)%,呈浓度依赖性诱导CNE2细胞凋亡(n=3,P<0.05)。200及400μmol/LDADS分别作用CNE2细胞24h后,DNA凝胶电泳出现典型梯形条带。Western blot检测显示,50、100、200和400μmol/LDADS处理CNE2细胞24h后,Bcl-2蛋白与β-Actin的灰度值比分别是1.92±0.21、1.21±0.18、0.89±0.14及0.41±0.09,与对照组(2.24±0.26)比较,差异有显著性(P<0.05);Bax蛋白与β-Actin的灰度值比分别是0.78±0.14、1.12±0.19、1.54±0.22及2.08±0.27,与对照组(0.33±0.08)比较,差异有显著性(P<0.05);随浓度的增加,Bcl-2蛋白下调,Bax蛋白表达上调。200、400μmol/LDADS处理CNE2细胞24h后,与对照组相比,Caspase3酶原蛋白条带变细并且可见到蛋白裂解产物,表明DADS可以促进Caspase3的活化。结论:DADS具有抑制人鼻咽癌CNE2细胞增殖和诱导凋亡的作用,其机制可能与Bcl-2蛋白下调,Bax蛋白表达上调导致Bcl-2/Bax比值下降,促进Caspase3的活化有关。Background and purpose： Diallyl disulfide （DADS） is an oil-soluble organosulfur compound found in garlic. It can induce apoptosis in various cancer cell lines in vitro. This study investigated the effect of apoptosis of nasopharyngeal cancer cells line CNE2 induced by diallyl disulfide （DADS） and its molecular mechanisms. Methods： The growth inhibition and apoptosis of CNE2 ceils and the protein levels of Bcl-2, Bax and Caspase 3 were examined by MTT assay, light microscopy, flow cytometry, DNA agarose gel electrophoresis and Western blot, respectively. Results： MTF assay showed that 50, 100, 200,400 μmol/L DADS significantly inhibited proliferation of CNE2 cells at 48 hr, its inhibition ratio were 3.66%, 14.28%, 29.66%, 61.28%, respectively, in dose-dependent manner （P 〈0.05）. After exposure to DADS, partial cells presented characteristic morphological changes of apoptosis under the light microscopy, Flow cytometry analysis showed that CNE2 cells treated with 50, 100, 200, 400μmol/L DADS for 24 hours significantly increased the percentageofapoptosiscells, its apoptosis rate were （10. 8 ±1. 3） %, （14.8±1.1）%, （21.7 ±2.0）%, （29.8 ±2.6）%, respectively （P 〈0.05）. ＂Ladder＂ bands occurred after exposure to 200, 400 μmol/L DADS for 24 hr in DNA agarose gel electrophoresis. Western blot showed that Bcl-2 expression were decreased and signal intensity for control group, ceils treated with 50, 10（3, 200, 400μmol/L DADS were （2.24 ±0.26）, 1.92 ±0.21, 1.21 ±0. 18, O. 89 ±0.14 and O. 41 ± 0.09, respectively. Bax expression were increased and signal intensity for control group, ceils treated with 50, 100, 200, 400μmol/L DADS were （0.33 ±0.08）, 0.78 ±0.14, 1.12 ±0.19, 1.54 ±0.22 and 2.08 ±0.27 respectively. CNE2cells treated with DADS at the concentration of 200, 400μmol, L^-1 for 24 hr, pro-Caspase 3 was decreased and cleaved product was detected. Conclusions： DADS could significantly inhibit proliferation and induce apoptosis of human nasopharyngea

关 键 词：二烯丙基二硫 鼻咽癌 CNE2细胞 凋亡 

分 类 号：R739.63[医药卫生—肿瘤]