5-aza-dC对293A细胞增殖的影响  

The effect of 5-aza-2’-deoxycytidine on the growth suppression of 293A cells

作　　者：刘璐[1] 张伟[2] 张颜波[3] 姜树原[1] 刘友[1] 黄丽华[1] 孟宪梅[2] 邵国[1] 

机构地区：[1]包头医学院生物医学研究中心及基础医学部,内蒙古包头014060 [2]包头医学院第二附属医院消化科,内蒙古包头014030 [3]泰山医学院附属医院神经内科,山东泰安271000

出　　处：《泰山医学院学报》2013年第6期401-404,共4页Journal of Taishan Medical College

基　　金：国家自然科学基金资助项目(81060212;81160244);内蒙古自然科学基金资助项目(2010BS1104);中国博士后基金项目(20080430851);内蒙古自治区高等学校科研项目(NJZY12221;NJZY12225)

摘　　要：目的观察5-氮-2'脱氧胞苷(5-aza-2'deoxycytidine,5-aza-dC)对293A细胞增殖的影响,初步探讨其在治疗肾脏肿瘤的作用及可能机制。方法实验组以1.0μmol/L的5-aza-dC处理人胚肾癌293A细胞,对照组加等体积的磷酸缓冲液(PBS),用MTT法测定实验组和对照组1～4 d的光密度值,通过比较实验组和对照组的第4天吸光度值来计算抑制率,流式细胞仪检测5-aza-dC对293A细胞株细胞周期的影响,实时定量PCR检测相关基因DNA甲基转移酶(DNMT)1、3A和3B mRNA的表达。结果 MTT检测实验组和对照组第4天的光密度分别为0.0663±0.0034和0.5453±0.0311,5-aza-dC能抑制人胚肾癌293A细胞的增殖;实验组与对照组相比,实验组91%细胞被阻滞在S期,5-aza-dC处理细胞后使细胞周期发生变化;实验组DNMT1、DNMT3A和DNMT3B mRNA相对于beta-actin mRNA的丰度值分别是0.036810892±0.007,0.008603032±0.001和0.023717786±0.005,而对照组相对丰度值是0.05594012±0.009,0.0362811±0.011和0.8751847±0.092;5-aza-dC处理使DNMT1、DN-MT3A和DNMT3B mRNA表达降低(P<0.05)。结论 5-aza-dC能抑制293A细胞株的增殖,其作用可能是通过降低DNMTs表达来干扰正常的细胞周期实现的。Objective： To study the growth suppression effect of 5-aza-2’-deoxycytidine on human embryo kidney 293A cells.Methods： There were two groups in this study： the 5-aza-dC（1.0μmol/L） group and the control group（add equal volume PBS）.The effect of 5-aza-dC on human embryo kidney 293A cells was measured using the MTT assay.The effect of 5-aza-dC on cell cycle was analyzed by flow cytometry analysis.The mRNA levels of DNMTs were analyzed by real-time PCR.Results： The absorb value of 5-aza-dC group and control group were 0.0663 ± 0.0034 and 0.5453 ± 0.0311 respectively in the 4th day by MTT assay.This result showed that 5-aza-dC inhibited the growth of 239A cell line;S-phase cell in 5-aza-dC treatment group was about 91% and it was very different from that in the control group.The DNMT1,DNMT3A and DNMT3B mRNA relative to beta-actin mRNA in 5-aza-dC group were 0.036810892 ± 0.007,0.008603032 ± 0.001 and 0.023717786 ± 0.005.They were 0.05594012 ± 0.009,0.0362811 ± 0.011 and 0.8751847 ± 0.092 in the control group.5-aza-dC down-regulated the mRNA expression of DNMTs.Conclusion： 5-aza-dC can suppress the growth of human embryo kidney 293A cells by decreasing the expression of DNMTs to interrupt the normal cell cycle.

关 键 词：5-氮-2'脱氧胞苷 293A细胞 增殖 

分 类 号：R737.11[医药卫生—肿瘤]