地西他滨对淋巴瘤细胞株Raji中P15INK4B基因的去甲基化作用

山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (7): 61-.

地西他滨对淋巴瘤细胞株Raji中P15INK4B基因的去甲基化作用

徐昕, 戴秋新, 徐茂忠, 赵钰, 孟文俊

东南大学医学院附属江阴医院血液科, 江苏江阴 214400

收稿日期:2010-04-16 出版日期:2010-07-16 发布日期:2010-07-16

Decitabine induces demethylation and up-regulates transcription of the P15INK4B gene in lymphoma cell line Raji cells

XU Xin, DAI Qiuxin, XU Maozhong, ZHAO Yu, MENG Wenjun

Department of Hematology, Affiliated Jiangyin Hospital of Medical College of Southeast University, Jiangyin 214400, Jiangsu, China

Received:2010-04-16 Online:2010-07-16 Published:2010-07-16

摘要/Abstract

摘要：

目的探讨Burkitt淋巴瘤细胞系Raji中P15INK4B基因甲基化状态，地西他滨对Raji细胞P15INK4B基因去甲基化作用及生长增殖的生物学影响。方法用不同浓度地西他滨作用淋巴瘤细胞株Raji，用台酚蓝拒染法研究药物对Raji细胞生长曲线的影响，采用流式细胞术检测细胞凋亡率，采用聚合酶链反应(RTPCR)检测P15INK4B表达，用甲基化特异PCR(MSP) 方法检测P15INK4B 基因的甲基化程度。结果地西他滨对Raji细胞有生长抑制作用，作用后细胞凋亡增加，P15INK4B基因表达增加，地西他滨使其甲基化程度下降。结论在Raji细胞中P15INK4B高度基因甲基化，并且表达下降，地西他滨通过去甲基化抑制淋巴瘤增殖。

关键词: 淋巴瘤；P15INK4B；地西他滨；甲基化

Abstract:

Objective To investigate methylation of the P15INK4B gene in lymphoma cell line Raji cells and to evaluate the effects of decitabine on demethylation of P15INK4B in human lymphoma cell line Raji cells and on proliferation of Raji cells. Methods Raji cells were treated with decitabine, and the effect of decitabine on proliferation of Raji cells wth trypan blue exclusion was investigated and the apoptosis of cells stained by Annexin /Ⅴ-FITC was determined. The mRNA expression of P15INK4B in cells treated with decitabine was determined by reverse transcription-polyerase chain reaction (RTPCR). Methylation of the P15INK4B gene in Raji cells was determined by PCR using methylation specific primer (MSP). Results Decitabine inhibited the Raji cell growth and promoted apoptosis of Raji cells, and up-regulated the P15INK4B mRNA expression by demethylation of the p15INK4B gene. Conclusion P15INK4B gene of Raji cells was methylated and down-regulated. Decitabine inhibited Raji cell growth by demethylation of P15INK4B.

Key words: Lymphoma； P15INK4B； Decitabine; Methylation

中图分类号:

R733

徐昕, 戴秋新, 徐茂忠, 赵钰, 孟文俊. 地西他滨对淋巴瘤细胞株Raji中P15INK4B基因的去甲基化作用[J]. 山东大学学报(医学版), 2010, 48(7): 61-.

XU Xin, DAI Qiuxin, XU Maozhong, ZHAO Yu, MENG Wenjun . Decitabine induces demethylation and up-regulates transcription of the P15INK4B gene in lymphoma cell line Raji cells[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2010, 48(7): 61-.

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