去甲基化剂5-氮杂-2’-脱氧胞苷对胃癌细胞生物学行为的影响  

Effect of the demethylating agent 5,Aza,2',deoxycytidine on the growth of human gastric carcinoma cell lines.

作　　者：杜鹏[1] 张一楚[1] 陈宏新[2] 姜宏华[1] 欧敬民[1] 钱惠琴[1] 

机构地区：[1]上海第二医科大学附属新华医院普外科,上海200092 [2]上海第二医科大学附属新华医院检验科,上海200092

出　　处：《外科理论与实践》2004年第3期217-220,共4页Journal of Surgery Concepts & Practice

摘　　要：目的:探讨去甲基化剂5氮杂2’脱氧胞苷(5AzaCdR)对胃癌细胞生物学行为的影响。方法:使用浓度5×106mol/L和105mol/L的5AzaCdR处理胃癌细胞株AGS、SGC7901、MKN28及MKN45。通过MTT、平板克隆实验观察处理前、后细胞的生长活性,RTPCR检测处理前、后抑癌基因RASSF1AmRNA表达的改变,并应用流式细胞仪进行细胞周期和凋亡率的分析。结果:4株胃癌细胞经不同浓度之5AzaCdR处理后,与对照组相比,生长速度出现不同程度减慢;实验组AGS细胞较对照组细胞克隆形成率显著降低(32.4%、28.5%比57.0%,P<0.01);5AzaCdR处理后,3株无RASSF1AmRNA表达的细胞(AGS、MKN28和SGC7901)均检出基因重新表达;处理前后4株胃癌细胞无明显G1期、G2/M期改变,AGS细胞凋亡率由0.6%增至18.6%和20.2%,MKN28细胞亦出现凋亡率由1.85%增至3.85%和6.61%。结论:去甲基化剂5AzaCdR对胃癌细胞生长周期无显著影响,可能系通过诱导抑癌基因再表达或凋亡等途径,间接抑制胃癌细胞的生长。Objective To explore the effect of demethylating agent 5,Aza,2',deoxycytidine on the growth of human gastric carcinoma cell lines. Methods Four gastric carcinoma cell lines (AGS, SGC7901, MKN28 and MKN45) were treated with 5×10,6M and 10,5M 5,Aza,CdR. Activation in cell growth was observed by MTT assay and colony formation on plate. The change in expression of the tumor suppressor gene RASSF1A mRNA was observed by using RT,PCR before and after 5,Aza,CdR treatment. The cell cycle and apoptosis were analyzed by flow cytometry. Results The 4 gastric carcinoma cell lines treated with two different doses of 5,Aza,CdR displayed a slowed growth rate in contrast to the control group. The colony formation rate of AGS cell treated with 5,Aza,CdR.(5×10,6 M,10,5 M) decreased dramatically (32.4%?28.5% vs 57.0%,P<0.01). The gene was reactivated by 5,Aza,CdR treatment in three of four cell lines not expressing RASSF1A. There were no G1 or G2/M phase arrests in the 4 cell lines treated with 5,Aza,CdR. In the AGS cell line, the apoptotic rate increased from 0.6% to 18.6% and 20.2%; and in MKN28, the apoptotic rate also increased from 1.85% to 3.85% and 6.61%. Conclusions The demethylating agent 5,Aza,CdR has no apparent influence on the cell cycle of gastric carcinoma cells. It may inhibit indirectly the growth of gastric carcinoma cells by inducing tumor suppressor genes re,expression or cell apoptosis.

关 键 词：胃癌细胞 5-氮杂-2’-脱氧胞苷 去甲基化 

分 类 号：R735.2[医药卫生—肿瘤]