摘要
本研究旨在探索多发性骨髓瘤细胞株U266 DNA甲基转移酶(DNMT)活性、基因表达情况并分析其生物学意义。用ELISA方法检测U266细胞DNMT活性,用半定量RT-PCR技术分析DNMT 1、3a、3b mRNA在U266细胞中的表达,体外用不同浓度去甲基化化合物异硫氰酸苯己酯(phenyl hexyleisothiocyanate,PHI)处理U266细胞,分析DNMT活性变化及DNMT1、3a、3b在瘤细胞中的表达改变。结果表明,骨髓瘤细胞U266中DNMT总活性升高,DNMT1、3a、3b mRNA均高表达,不同浓度PHI处理U266细胞后,细胞增殖抑制,DNMT活性亦被显著抑制,DNMT1、DNMT3a以及DNMT3b mRNA表达下降并呈浓度依赖性。结论:DNMT活性及其表达增高是骨髓瘤细胞特征之一,当DNMT活性或mRNA表达抑制时瘤细胞发生凋亡,因此DNMT可作为骨髓瘤治疗的新靶点。
In order to study the activity and gene expression of DNA methyltransferase（DNMT） on U266 myeloma cells and to analyze their significance,the activity of DNMT was detected by ELISA,and the expressions of DNMT1,DNMT3a and 3b were analyzed by RT-PCR.U266 cells were treated by phenylhexyleisothiocyanate（PHI）,and the change of activity and gene expression of DNMT were determined.The results indicated that the activity and expression of DNMT in U266 myeloma cells were higher,compared with normal control.After being treated by different concentration of PHI,U266 cells were driven into apoptosis and the activity of DNMT decreased obviously and the mRNA level of DNMT declined.It is concluded that the activity and gene expression of DNMT on U266 myeloma cells are higher,and DNMT may be a new therapeutic target of multiple myeloma.
出处
《中国实验血液学杂志》
CAS
CSCD
2011年第6期1429-1431,共3页
Journal of Experimental Hematology
基金
2010年厦门市科技计划项目
编号3502Z20104034
