5-氮杂-2′-脱氧胞苷对人肺腺癌裸鼠种植瘤的抑制作用及组织因子途径抑制物2基因甲基化状态和表达的影响  

5-aza-2′-deoxycytidine suppresses the growth of human lung adenocarcinoma cells in nude mouse xenograft models and its effect on methylation status and expression of TFPI-2 gene

作　　者：梁江水[1] 殷桂林[2] 董永强[2] 马忠厦[1] 肖跃华[1] 纪涛[2] 

机构地区：[1]郴州市第一人民医院胸部肿瘤肺癌诊疗中心,423000 [2]广州军区武汉总医院心胸外科

出　　处：《中华肿瘤杂志》2015年第7期485-489,共5页Chinese Journal of Oncology

基　　金：武汉市青年科技晨光计划项目（201050231090）

摘　　要：目的探讨去甲基化药物5-氮杂-2′-脱氧胞苷（5-Aza-CdR）对人肺腺癌裸鼠种植瘤生长的抑制作用，以及对种植瘤组织中组织因子途径抑制物2（TFPI-2）基因甲基化状态和表达的影响。方法建立人肺腺癌A549细胞裸鼠种植瘤模型，按5-Aza—CdR不同剂量分为对照组（未注射5-Aza—CdR）和实验组，其中实验组分为0．5mg／kg组、1mg／kg组和2mg／kg组。比较各组裸鼠种植瘤生长的变化。采用甲基化特异性聚合酶链反应、实时定量荧光聚合酶链反应和Western blot检测5-Aza-CdR对裸鼠种植瘤组织TFPI-2基因甲基化状态、mRNA和蛋白表达的影响。结果用药28d后处死裸鼠时，0．5mg／kg组、1mg／kg组、2mg／kg组和对照组荷瘤裸鼠体重分别为（26．80±0．18）g、（26．67±0．28）g、（26．50±0．26）g和（27．12±0．38）g，差异无统计学意义（P〉0．05）。0．5mg／kg组、1mg／kg组、2mg／kg组和对照组皮下种植瘤体积分别为（400．67±2．68）mm^3、（285．71±2．91）mm^3、（230．44±3．15）mm^3和（709．22±2．87）mm^3，差异有统计学意义（P〈0．05）。对照组TFPI-2基因呈完全甲基化状态，0．5mg／kg组和1mg／kg组呈不完全甲基化状态，2mg／kg组呈非甲基化状态。0．5mg／kg组、1mg／kg组、2mg／kg组和对照组TFPI-2 mRNA的相对表达水平分别为1．67±0．07、3．40±0．24、5．55±0．61和1．00±0．00，差异有统计学意义（P〈0．05）。0．5mg／kg组、1mg／kg组、2mg／kg组和对照组TFPI-2蛋白表达的相对水平分别为0．36±0．01、0．64±0．02、0．81±0．20和0．18±0．02，差异有统计学意义（P〈0．05）。结论5-Aza-CdR能抑制人肺腺癌裸鼠种植瘤的生长，诱导种植瘤细胞TFPI-2基因的表达，其机制可能为5-Aza-CdR通过去甲基化作用使甲基化的TFPI-2基因重新表达，从而抑制人肺腺癌A549细胞的生长和增殖。Objective To investigate the inhibitory effect of classic demethylating drug 5-aza-2′- deoxycytidine（5-Aza-CdR） on the growth of human lung adenocarcinoma cells in nude mouse xenograft models, and to observe its effect on methylation status and expression of TFPI-2 gene in the nude mouse xenograft tissues. Methods The nude mouse xenograft model was established by subcutaneous inoculation of human lung adenocarcinoma A549 cells.According to different doses of 5-Aza-CdR, the tumor-bearing nude mice were randomly divided into experimental groups （0.5 mg/kg group, 1 mg/kg group, 2 mg/kg group） and control group （0 mg/kg group）. The tumor growth in the nude mice was observed. The methylation status and the expression of TFPI-2 gene mRNA and protein were detected by methylation specific potymerase chain reaction, real-time fluorescent quantitative polymerase chain reaction and Western blot assay. Results The nude mice were euthanized at 28 days after intraperitoneal injection of 5-Aza-CdR. The body weight of tumor-bearing nude mice was （27.12±0.38）g in the 0 mg/kg group, （26.80±0.18）g in the 0.5 mg/kg group, （26.67±0.28）g in the 1 mg/kg group, and （26.50±0.26）g in the 2 mg/kg group, showing no significant difference among them （P〉0.05）. The volume of xenograft tumors in the 0 mg/kg group was （709.22±2.87）mm^3, （400.67±2.68） mm^3 in the 0.5 mg/kg group, （285.71±2.91）mm^3 in the 1 mg/kg group, and （230.44±3.15）mm^3 in the 2 mg/kg group, showing a significant difference （P〈0.05）. There were complete methylation of TFPI-2 gene in the 0 mg/kg group, incomplete methylation in the 0.5 and 1 mg/kg groups, and unmethylation in the 2 mg/kg group. The relative mRNA level in the 0, 0.5, 1, 2 mg/kg groups were 1.00±0.00, 1.67±0.07, 3.40±0.24, and 5.55±0.61, respectively （P〈0.05）. The relative expression level of TFPI-2 protein in the 0, 0.5, 1, 2 mg/kg groups was 0.18±0.02, 0.36±0.01, 0.64±0.02, and 0.81±0.20, respectively （P〈0.05）. Conclusions 5-

关 键 词：肺肿瘤 腺癌 小鼠 DNA甲基化 5-氮杂-2′-脱氧胞苷 组织因子途径抑 制物2 

分 类 号：R734.2[医药卫生—肿瘤]