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

作　　者：郭文娴 张敏[2] 孙立荣[1] 雷珂[3] 王红芳 宋爱琴[1] 

机构地区：[1]青岛大学附属医院血液儿科,山东青岛266003 [2]济宁医学院附属医院儿科,山东济宁272000 [3]青岛大学附属医院儿科研究所,山东青岛266000 [4]青岛市妇女儿童医院血液科,山东青岛266034

出　　处：《现代生物医学进展》2016年第27期5232-5236,共5页Progress in Modern Biomedicine

基　　金：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)

摘　　要：目的:探讨5-氮杂-2'-脱氧胞苷(5'-Aza-Cd R)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-Cd R(1μg/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 m RNA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 m RNA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-Cd R能抑制人伯基特淋巴瘤的生长,诱导瘤细胞CHFR m RNA表达增加,其机制可能为5'-Aza-Cd R使甲基化的CHFR启动子发生去甲基化。Objective： To determine the inhibitory effect of 5’-Ara-Cd R 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-Cd R（1 μg/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 m RNA 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 m RNA 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-Cd R can inhibit the growth of Burkitt Lymphoma, increase the expression of CHFR m R- NA in cancer cells.The mechanism may be that 5’-Aza-Cd R makes the methylated CHFR demethylate.

关 键 词：淋巴瘤 RAJI细胞 CHFR基因 5-氮杂-2’-脱氧胞苷 DNA甲基化 

分 类 号：R733.4[医药卫生—肿瘤]