紫杉醇诱导人乳腺癌细胞凋亡过程DNA甲基化水平检测  

DETECTION OF CELLULAR DNA METHYLATION IN TAXOL INDUCED APOPTOSIS OF HUMAN BREAST CANCER CELLS

作　　者：陈于法[1,2,3] 郑树[1,2,3] 陈丽荣 李士敏[1,2,3] 

机构地区：[1]浙江医科大学肿瘤研究所 [2]浙江医科大学分析中心 [3]杭州九源基因工程有限公司

出　　处：《肿瘤》1999年第2期65-68,共4页Tumor

摘　　要：目的紫杉醇诱导细胞凋亡过程中ＤＮＡ甲基化水平状态及其意义，尚未见报道。方法本文应用反相高效液相色谱法和琼脂糖凝胶电泳法对紫杉醇诱导人乳腺癌细胞（ＢＣａｐ３７）凋亡过程中ＤＮＡ甲基化水平进行分析，并与秋水仙碱、顺铂、鬼臼乙叉甙、阿霉素和放线菌酮处理组进行比较。结果紫杉醇处理ＢＣａｐ３７细胞３小时，ＤＮＡ甲基化水平降低；在处理１２～２４和４８小时时，ＤＮＡ甲基化水平逐渐上升，在４８小时达到最高峰，显著高于对照组（Ｐ＜０．０２）。然而在处理７２小时，ＤＮＡ甲基化水平明显降低，并低于对照组（Ｐ＜０．０５）。除阿霉素处理ＢＣａｐ３７细胞能够使ＤＮＡ甲基化水平降低之外，其余抗癌药处理组细胞ＤＮＡ甲基化水平均有轻度的增高。结论在紫杉醇诱导乳腺癌细胞凋亡过程中，细胞ＤＮＡ甲基化水平增高可能与先前研究发现的腺苷基蛋氨酸合成酶基因表达增高密切相关。本文就ＤＮＡ高甲基化反应在紫杉醇诱导的细胞凋亡中的潜在生物学意义进行了讨论。Objective:Cellular DNA methylation status and biological functon in the process of taxol induced apoptosis remains unclear.Methods:Reversed phase high performance liquid chromatography and 1% agarose gel electrophoresis were utilized in the detection of DNA methylation level and DNA fragmentation in taxol induced apoptosis of human breast cancer cells(BCap37). The DNA methylation level resulted from taxol treatment was compared with that from colchicine,cis platin,etoposide,doxorubicin and cycloheximide treatments respectively.Results:Taxol treatment at 3 hours resulted in temporary decrease of DNA methylation.Then,the hypermethylation of DNA gradually appeared at 12,24 and 48 hours of taxol treatment.At 48 hours,the DNA methylation reaction reached a peak,which was significantly higher than that of control group( P <0 02) However,at 72 hours,when apoptotic cell death became evident ,the DNA methylation level was reduced significantly lower than that of the control( P <0 05).Except for doxorubicin resulting in the decrease of DNA methylation,colchicine,cis platin,etoposide or cycloheximide treatment slightly increased the DNA methylation level.Conclusion:In the taxol induced apoptosis the hypermethylation of cellular DNA is closely associated with the up regulation of S adenosylmethionine synthetase gene previously identified.The potential biological significance of the hypermethylation of cellular DNA in taxol induced apoptosis was discussed.

关 键 词：细胞凋亡 紫杉醇 DNA甲基化 乳腺肿瘤 细胞株 

分 类 号：R737.902[医药卫生—肿瘤]