白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2012年
4期
213-217
,共5页
周霞%张鹏%周翔%孟月生
週霞%張鵬%週翔%孟月生
주하%장붕%주상%맹월생
淋巴瘤,膜细胞%p151NK4B%p27kip1%甲基化%细胞凋亡%地西他滨
淋巴瘤,膜細胞%p151NK4B%p27kip1%甲基化%細胞凋亡%地西他濱
림파류,막세포%p151NK4B%p27kip1%갑기화%세포조망%지서타빈
Lymphoma,mantle-cell%p151NK4B%p27kipl%Methylation%Apoptosis%Decitabine
目的 探讨套细胞淋巴瘤细胞系Mino细胞中p15INK4B和p27kip1基因甲基化状态,以及地西他滨对Mino细胞p15INK4B和p27kip1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Western blot检测p15INK4B、p27kip1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p27kip1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G1期,p15INK4B和p27kip1基因mRNA表达增加,而抗凋亡基因bcl-2 mRNA的表达下降.经6.4、3.2、1.6 mmol/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％,p27kip1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p27kip1基因启动子高度甲基化,并且p15INK4B及p27kip1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p27kip1基因去甲基化作用及对bcl-2基因表达的调控所致.
目的 探討套細胞淋巴瘤細胞繫Mino細胞中p15INK4B和p27kip1基因甲基化狀態,以及地西他濱對Mino細胞p15INK4B和p27kip1基因去甲基化及誘導細胞凋亡的作用及機製.方法 用不同濃度地西他濱處理套細胞淋巴瘤細胞繫Mino細胞,用錐蟲藍拒染法研究藥物作用後細胞生長情況,繪製生長麯線,採用流式細胞術分析細胞凋亡和細胞週期的變化,採用RT-PCR和Western blot檢測p15INK4B、p27kip1及bcl-2基因mRNA和蛋白的錶達水平,用甲基化特異PCR方法檢測p15INK4B及p27kip1基因的甲基化程度.結果 地西他濱對套細胞淋巴瘤細胞繫Mino細胞有生長抑製作用,作用後細胞凋亡增加,細胞週期阻滯在G1期,p15INK4B和p27kip1基因mRNA錶達增加,而抗凋亡基因bcl-2 mRNA的錶達下降.經6.4、3.2、1.6 mmol/L地西他濱作用72h後,Mino細胞p15INK4B基因啟動子甲基化率分彆下降至25.2％、48.2％和65.0％,p27kip1基因啟動子甲基化率分彆下降至20.2％、50.2％和70.0％.結論 在套細胞淋巴瘤細胞繫Mino細胞中存在p15INK4B及p27kip1基因啟動子高度甲基化,併且p15INK4B及p27kip1基因mRNA錶達水平下降.地西他濱誘導Mino細胞凋亡,可能是通過對p15INK4B及p27kip1基因去甲基化作用及對bcl-2基因錶達的調控所緻.
목적 탐토투세포림파류세포계Mino세포중p15INK4B화p27kip1기인갑기화상태,이급지서타빈대Mino세포p15INK4B화p27kip1기인거갑기화급유도세포조망적작용급궤제.방법 용불동농도지서타빈처리투세포림파류세포계Mino세포,용추충람거염법연구약물작용후세포생장정황,회제생장곡선,채용류식세포술분석세포조망화세포주기적변화,채용RT-PCR화Western blot검측p15INK4B、p27kip1급bcl-2기인mRNA화단백적표체수평,용갑기화특이PCR방법검측p15INK4B급p27kip1기인적갑기화정도.결과 지서타빈대투세포림파류세포계Mino세포유생장억제작용,작용후세포조망증가,세포주기조체재G1기,p15INK4B화p27kip1기인mRNA표체증가,이항조망기인bcl-2 mRNA적표체하강.경6.4、3.2、1.6 mmol/L지서타빈작용72h후,Mino세포p15INK4B기인계동자갑기화솔분별하강지25.2％、48.2％화65.0％,p27kip1기인계동자갑기화솔분별하강지20.2％、50.2％화70.0％.결론 재투세포림파류세포계Mino세포중존재p15INK4B급p27kip1기인계동자고도갑기화,병차p15INK4B급p27kip1기인mRNA표체수평하강.지서타빈유도Mino세포조망,가능시통과대p15INK4B급p27kip1기인거갑기화작용급대bcl-2기인표체적조공소치.
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 p15INK4B and p27kip1 genes and on apoptosis of Mino cells and its relative mechanism. Methods Mino cells were after treated with various concentration of decitabine, 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 p15INK4B 、p27kip1 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 p15INK4B 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 p15INK4B gene were decreased to 25.2 ％ 、48.2 ％ and 65.0 ％respectively,in the meantime,the methylationg of p27kip1 gene was decreased to 20.21％ 、50.2 ％ and 70.0 ％. Conclusion The hp15INK4B and p27kip1 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 bcl-2 gene and demethylation of p15INK4B and p27kip1 genes.
