论文部分内容阅读
目的探究小儿肾母细胞瘤(WT)组织和血液中SIX2基因在mRNA水平的相对表达量及启动子是否发生甲基化,并观察其与患儿的基本临床资料之间的关系,以期为临床靶向基因治疗提供科学的依据。方法选择2011年3月至2015年7月就诊并确诊的64例WT患儿[男30例,女34例;年龄3~67(26.9±8.6)月]为研究对象,运用实时荧光定量PCR(qRT-PCR)和甲基化特异性聚合酶链反应(MSP)分别检测患儿WT组织、瘤旁组织和外周血以及18例健康儿童外周血中SIX2基因在mRNA水平的相对表达量及启动子甲基化状态。结果 WT患儿瘤组织SIX2 mRNA的相对表达量明显高于瘤旁正常组织(P<0.01),SIX2 mRNA启动子的甲基化阳性率明显低于瘤旁正常组织(P<0.01);WT患儿外周血中SIX2 mRNA的表达量明显高于对照组外周血(P<0.01),SIX2基因启动子的甲基化阳性率明显低于对照组外周血(P<0.01)。WT患儿组织SIX2基因相对表达量与患儿肿瘤大小、临床分期、病理类型、淋巴转移、血管浸润有关(P<0.05,P<0.01),而与性别、年龄和复发无明显关系(P均>0.05)。SIX2基因启动子的甲基化与上述临床资料均无明显关系(P均>0.05)。随访36个月,死亡42例,存活22例,3年存活率34.4%;SIX2 mRNA相对表达量>2.8和≤2.8的患儿存活率(23.8%vs 54.5%,P<0.05)和存活时间[(13.3±5.2)月vs(22.2±8.7)月,P<0.01]比较,差异均有统计学意义。SIX2启动子甲基化表达阴性和阳性的患儿3年存活率(14.3%vs 44.2%,P<0.05)和存活时间[(14.5±4.5)月vs(22.1±4.9)月,P<0.01]比较,差异均有统计学意义。结论 SIX2基因的高表达和其启动子的未甲基化可能是WT的发病基础,SIX2基因或可作为WT基因治疗的靶目标。
Objective To investigate the mRNA expression of SIX2 in pediatric nephroblastoma (WT) and blood and the promoter methylation, and to observe its relationship with the basic clinical data of children, with a view to clinical application Targeted gene therapy to provide a scientific basis. Methods Sixty-four cases of WT children (30 males and 34 females, aged 3-67 (26.9 ± 8.6) months) from March 2011 to July 2015 were selected as study subjects. Real-time quantitative PCR qRT-PCR) and methylation-specific polymerase chain reaction (MSP) were used to detect the relative expression level of SIX2 mRNA in peripheral blood of WT tissues, peritumoral tissues and peripheral blood of 18 children and healthy children Methylation status. Results The relative expression of SIX2 mRNA in WT tissues was significantly higher than that in normal tissues (P <0.01). The positive rate of SIX2 mRNA promoter was significantly lower than that in normal tissues (P <0.01) The expression of SIX2 mRNA in peripheral blood of children was significantly higher than that of the control group (P <0.01). The methylation positive rate of SIX2 promoter was significantly lower than that of the control group (P <0.01). The relative expression of SIX2 in WT was correlated with tumor size, clinical stage, pathological type, lymph node metastasis and vascular infiltration (P <0.05, P <0.01), but not with gender, age and relapse (P > 0.05). Methylation of SIX2 gene promoter had no significant relationship with the above clinical data (all P> 0.05). The survival rate was 23.8% vs 54.5% (P <0.05) and the survival time was significantly higher in children with SIX2 mRNA expression> 2.8 and ≤2.8 after 36 months of follow-up (42 deaths, 22 survivors and 34.4% (13.3 ± 5.2) month vs (22.2 ± 8.7) months, P <0.01], the differences were statistically significant. The 3-year survival rate (14.3% vs 44.2%, P <0.05) and survival time in the SIX2-negative and -positive methylation-expressing children [(14.5 ± 4.5) months vs (22.1 ± 4.9) months, P <0.01] Comparison, the differences were statistically significant. Conclusion The high expression of SIX2 gene and the unmethylation of its promoter may be the basis for the pathogenesis of WT. The SIX2 gene may serve as the target of WT gene therapy.