地西他滨对套细胞淋巴瘤Mino细胞系的作用及其机制  

Effect and related mechanism of decitabine on mantle cell lymphoma cell line Mino

作　　者：周霞[1] 张鹏[1] 周翔[1] 孟月生[1] 

机构地区：[1]青岛市市立医院血液科,266011

出　　处：《白血病．淋巴瘤》2012年第4期213-217,共5页Journal of Leukemia & Lymphoma

摘　　要：目的探讨套细胞淋巴瘤细胞系Mino细胞中p15INK4B和p27kipl基因甲基化状态，以及地西他滨对Mino细胞p15INK4B和p27kipl基因去甲基化及诱导细胞凋亡的作用及机制。方法用不同浓度地西他滨处理套细胞淋巴瘤细胞系Mino细胞，用锥虫蓝拒染法研究药物作用后细胞生长情况，绘制生长曲线，采用流式细胞术分析细胞凋亡和细胞周期的变化，采用RT-PCR和Westernblot检测D15INK4B、p27kilc-1及bcl-2基因mRNA和蛋白的表达水平，用甲基化特异PCR方法检测p15INK4B及D27kipl基因的甲基化程度。结果地西他滨对套细胞淋巴瘤细胞系Mino细胞有生长抑制作用，作用后细胞凋亡增加，细胞周期阻滞在G，期，p15INK4B和p27kipl基因mRNA表达增加，而抗凋亡基因bcl-2mRNA的表达下降。经6．4、3．2、1．6mmol／L地西他滨作用72h后，Mino细胞p15INK4B基因启动子甲基化率分别下降至25．2％、48．2％和65．0％，p27kipl基因启动子甲基化率分别下降至20．2％、50．2％和70．0％。结论在套细胞淋巴瘤细胞系Mino细胞中存在p15INK4B及p27kipl基因启动子高度甲基化，并且p15INK4B及p27kipl基因mRNA表达水平下降。地西他滨诱导Mino细胞凋亡，可能是通过对p15INK4B及p27kipl基因去甲基化作用及对bcl-2基因表达的调控所致。Objective To investigate methylation of the P15INK4B and p27kipl genes in human mantle cell lymphoma cell line Mino,to evaluate the effects of decitabine on demethylation of pl5INK4B and p27kipl genes and on apoptosis of Mino cells and its relative mechanism. Methods Mino cells were after treated with various concentration of deeitabine, the cell viability, cell cycle distribution or the apoptosis of Mino cells was respectively analyzed by trypan dye-exclusion assay or flow cytometry. The mRNA and protein expression of pl51NK4B ,p27kipl and bcl-2 were studied by RT-PCR or Western blot, respectively . Methylation of the p15INK4B and p27kip1 genes in Mino cells were determined by PCR using the methylation specific primer（MSP）.Results Decitabine significantly inhibit the cell growth,induced G1 arrest and promoted apoptosis of Mino cells . The expression of pl5INK4B and p27kipl mRNA were both significantly increased ,wheres bcl-2 mRNA was decreased. After treatment with 6.4,3.2,1.6 mmol/L decitabine for 72 h,the methylationg of pl5INK4B gene were decreased to 25.2 % ,48.2 % and 65.0 % respectively,in the meantime,the methylationg of p27kipl gene was decreased to 20.21% ~50.2 % and 70.0 %o Conclusion The hpl5INK4B and p27kipl genes of Mino cells are methylated and down-regulated. Decitabine can inhibit the proliferation and induce the apoptosis of Mino cell lines,with the reduction of bel-2 gene and demethylation of pl5INK4B and p27kipl genes.

关 键 词：淋巴瘤 膜细胞 p151NK4B P27KIP1 甲基化 细胞凋亡 地西他滨 

分 类 号：R733.1[医药卫生—肿瘤] R965[医药卫生—临床医学]