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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 investigate the inhibitory effect of 5-aza-2’-deoxycytidine (5’-Aza-Cd R) on human Burkitt’s lymphoma Raji xenografts and its effect on the expression of CHFR and methylation Impact. Methods: 18 nude mice were inoculated subcutaneously with Raji cell suspension to establish 18 nude mice models and randomly divided into experimental group and control group. The cells were injected intraperitoneally with equal volume of 5’-Aza-Cd R (1μg / g) and physiological Saline, calculate the size of the tumor volume, draw the tumor growth curve, and compare the size and weight of the two groups of tumor size after 24 days. 2 DNA and RNA were extracted from xenograft tumor tissues respectively. The expression of CHFR m RNA was detected by real-time fluorescence quantitative PCR. The methylation status of CHFR gene was detected by methylation-specific polymerase chain reaction (MSP). Results: 1 The experimental group grew more slowly. At the end of the experiment, there was significant difference between the experimental group and the control group in tumor volume and weight (P <0.01). The expression of CHFR m RNA in experimental group was significantly higher than that in control group (3.69 ± 1.20 vs 1.04 ± 0.30, t = 3.94, P <0.001). 3 The methylation bands in the experimental group were significantly darker than those in the control group, and the unmethylated bands appeared at the same time. Only the methylated bands of the CHFR gene promoter in the control group were amplified. CONCLUSION: 5’-Aza-Cd R can inhibit the growth of human Burkitt’s lymphoma and induce the expression of CHFR m RNA in tumor cells. The possible mechanism is that 5’-Aza-Cd R causes methylated CHFR promoter to occur Methylation.