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郭文娴 张敏 孙立荣 雷珂 王红芳 宋爱琴.5'-Aza-CdR 对B细胞淋巴瘤的抑制作用及对瘤细胞CHFR 表达和甲基化的影响[J].现代生物医学进展英文版,2016,16(27):5232-5236.
5'-Aza-CdR 对B细胞淋巴瘤的抑制作用及对瘤细胞CHFR 表达和甲基化的影响
Inhibitory Effect of 5'-Ara-CdR on B-cell Lymphoma and Effects of it on the Expression and Methylation of CHFR in Tumor Cells
  
DOI:
中文关键词: 淋巴瘤  Raji 细胞  CHFR 基因  5-氮杂-2'-脱氧胞苷  DNA甲基化
英文关键词: Lymphoma  Raji cells  CHFR gene  5'-Ara-CdR  DNA methylation
基金项目:山东省自然科学基金项目(Y2008C170);青岛市自然科学基金项目(2012-1-3-2-(5)- nsh)
Author NameAffiliation
郭文娴 张敏 孙立荣 雷珂 王红芳 宋爱琴 青岛大学附属医院血液儿科济宁医学院附属医院儿科青岛大学附属医院儿科研究所青岛市妇女儿童医院血液科 
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中文摘要:
      目的:探讨5- 氮杂-2'-脱氧胞苷(5'-Aza-CdR)对人伯基特淋巴瘤Raji 裸鼠移植瘤的抑制作用,以及对瘤组织中CHFR表达 和甲基化的影响。方法:①皮下接种Raji细胞悬液构建荷瘤裸鼠模型18 只,并随机分为实验组和对照组,分别予腹腔内注射等体 积的5'-Aza-CdR(1 ug/g)及生理盐水,计算肿瘤体积大小,绘制肿瘤生长曲线,24天后比较两组移植瘤体积大小和重量。②分别从 移植瘤组织中提取DNA 及RNA,应用实时荧光定量PCR 检测CHFR mRNA 的表达,甲基化特异性聚合酶链反应(MSP)检测 CHFR 基因甲基化情况。结果:①实验组移植瘤生长速度较缓慢。实验结束时,实验组移植瘤体积、重量与对照组相比,差异有显 著统计学意义(P<0.01)。②实验组瘤细胞中CHFR mRNA的表达水平(3.69± 1.20)与对照组(1.04± 0.30)相比,差异有显著统计 学意义(t=3.94,P<0.001)。③实验组中的甲基化条带与对照组相比明显变暗,同时出现非甲基化条带,对照组中CHFR 基因启动 子只有甲基化条带被扩增。结论:5'-Aza-CdR能抑制人伯基特淋巴瘤的生长,诱导瘤细胞CHFR mRNA 表达增加,其机制可能为 5'-Aza-CdR 使甲基化的CHFR 启动子发生去甲基化。
英文摘要:
      Objective:To determine the inhibitory effect of 5'-Ara-CdR on the growth of Burkitt Lymphoma's xenografts in nude mice, to observe its effect on CHFR gene expression and methylation in the xenografts, and to explore the possible mechanisms.Methods:Raji cells were inoculated subcutaneously in 18 nude mice to establish xenograft models of Burkitt Lymphoma. Xenografts models were divided into experimental group and control group. Were intraperitoneally injected an equal volume of 5'-Aza-CdR (1 滋g/g) and saline, calculating the tumor size, drawing the tumor growth curve, All the mice were killed after 24 days observation end, stripping out the xenografts, which were compared tumor volume and weight. RNA and DNA was extracted from tumor tissue, applied the real-time quantitative PCR for the detection of the expression of CHFR mRNA in the two groups of transplanted tumor tissue and applied the methyl of specific polymerase chain reaction method (MSP) for the detection of the methylation status of CHFR gene in the two groups of transplanted tumor tissue.Results:①The tumor growth was slow in the expreimental group. At the end of the experiment, compared with the contral group, the tumor volume and weight of transplantation tumor in the experimental group the differences were statistically significant (P<0.01). ② Compared with the control group (1.04 ± 0.30), the expression level of CHFR mRNA in the experimental group (3.69± 1.20) was significantly increased, and the difference was significant (P<0.01). ③In the control group only methylation stripe of CHFR gene promoter were amplified by PCR. Methylation stripe in experimental group compared with control group obviously dim, and had the non methylation stripe.Conclusion:5'-Aza-CdR can inhibit the growth of Burkitt Lymphoma, increase the expression of CHFR mR- NA in cancer cells.The mechanismmay be that 5'-Aza-CdR makes the methylated CHFR demethylate.
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