二烯丙基二硫对人结肠癌HT-29细胞G_1期的阻滞作用  

Effect of diallyl disulfide on G_1 arrest in human colon cancer cell

作　　者：黄幼生[1] 苏琦[1] 周秀田[1] 苏坚[1] 廖前进[1] 解娜[1] 葛玲[1] 

机构地区：[1]南华大学肿瘤研究所,湖南衡阳421001

出　　处：《中国药理学通报》2005年第9期1128-1132,共5页Chinese Pharmacological Bulletin

摘　　要：目的探讨二烯丙基二硫(DADS)对人结肠癌HT-29细胞G1期的阻滞作用及分子机制。方法采用MTT、细胞计数法、流式细胞术和免疫细胞化学方法分析DADS对体外培养的HT-29细胞增殖抑制作用、细胞周期分布及细胞周期相关蛋白表达的影响。结果MTT法显示,60、120μmol.L-1DADS作用HT-29细胞24 h后,生长抑制率分别达23.1%、45.6%。细胞计数法表明,常规培养HT-29细胞群体倍增时间为22.58 h,60μmol.L-1DADS作用HT-29细胞后,其细胞群体倍增时间延长到31.20 h。流式细胞仪分析结果显示,60μmol.L-1DADS阻滞HT-29细胞在G1期,与对照组相比,可使G1期细胞增加约2倍,而120μmol.L-1DADS显著地将细胞阻滞在G2/M期。免疫细胞化学分析表明在G1期阻滞同时有p21W af1蛋白表达上调,Cyc lin E、C-myc蛋白表达下降。结论低剂量DADS对HT-29细胞的抑制增殖作用可能与G1期阻滞有关,DADS对HT-29细胞G1期阻滞的分子机制可能与调节p21W af1、Cyc lin E、C-myc表达相关。Aim To explore cell cycle of HT-29 cells. the effects of DADS in the Methods HT-29 cells growth inhibition were measured by MTI＇ assay and cell counting. Phase distribution of cell cycle was analyzed by flow cytometry. Expression of p21 Wall, C-myc, Cyc- lin E was determined by immunohisto- chemistry analysis. Results MTT assay showed that DADS significantly inhibited HT-29 cells and exhibited a time, dose- dependent model. Adding 60μmol·L^-1 and 120μmol·L^-1 DADS for 24 h suppressed HT-29 growth by 23.1% ,45.6% respectively. Cell counting showed that average doubling time retarded from 22.58 h in normal cultured HT-29 cells to 31.2 h in 60umol·L^-1 DADS experimented HT-29 cells （ P 〈 0.05 ）. Flow cytometry analysis revealed that treating HT-29 cells with 60μmol·L^-1 DADS accumulated cells inthe G1 phase,however, 120 μmol·L^-1 DADS caused G2/M arrest. The proportion of cells in the G1 phase after treatment with 60μmol·L^-1 DADS for 24 h was 45.2 % , almost 2-folds that occurring in untreated cells （ 27.6% ）. Immunohistochemistry analysis revealed that 60μmol·L^-1 DADS increased the amount associated with p21 wan and decreased the level of cmyc, cyclin-E protein. Conclusion The antiproliferation property of 60μmol·L^-1 diallyl disulfide （DADS） in cultured human colon carcinoma HT-29 cell line relates to its ability to arrest G,. G, arrest of HT-29 cells by DADS may be related to the expression of p21 wan, C-myc, Cyclin-E protein.

关 键 词：二烯丙基二硫 结肠癌 HT-29细胞 细胞周期 eye-lin E蛋白 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R341[医药卫生—基础医学]