基于Keap1基因多态性的肾透明细胞癌分子标志物的研究

目的:探讨KeapJ基因单核苷酸多态性与肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)的相关性,为临床诊疗提供有效的分子靶点.方法:采集ccRCC组织108例与正常对照肾组织86例,采用聚合酶链式反应-限制性片段长度多态性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)和基因测序技术检测KeapJ基因单核苷酸多态性,判读两组样本基因型,实时荧光定量PCR法检测Keap1基因在两组中的表达情况.结果:Keap1基因rs1048289的不同基因型与等位基因频率在肿瘤组和对照组之间的分布不同,差异均有统计学意义(P＜0.05).AA基因型及A等位基因携带者患肾透明细胞癌的风险明显升高,AA基因型(OR=2.292,95％ CI:1.159～4.533,P＜0.05)及A等位基因(OR =2.067,95％ CI:1.280 ～3.342,P=0.001)为肾透明细胞癌患病的危险因素.肿瘤组Keap1基因表达量比对照组明显降低(P＜0.05),但不同基因型样本间Keap1基因表达量无明显差异(P＞0.05).结论:Keap1基因rs1048289与肾透明细胞癌患病显著相关,有望成为肾透明细胞癌早期诊断与基因治疗的有效分子靶点.

Research on the molecular markers of clear cell renal cell carcinoma based on polymorphisms of the Keap1 gene

Objective:To explore the correlation between single nucleotide polymorphism (SNP) of Keap1 gene and clear cell renal cell carcinoma (ccRCC),and to provide an effective molecular target for the clinical diagnosis and treatment of the ccRCC.Methods:One hundred and eight carcinoma tissues from the patients with ccRCC (tumor group) and 86 normal renal tissues from the tumor-free individuals (control group) were collected.Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and gene sequencing assays were used to detect NSP of the Keap1 gene and genotype of the samples of the both groups.Expression status of the Keap1 gene in the both groups were examined by real-time fluorescence quantitative PCR.Results:Distribution frequencies of the Keap1 gene rs1048289 in different genotypes and alleles were statistically different between the tumor group and the control group (P ＜ 0.05).Risk of ccRCC in the patients carraying AA genotype and A allele obviously increased,and AA genotype (OR =2.292,95％ CI:1.159-4.533,P ＜ 0.05) and A allele (OR =2.067,95％ CI:1.280-3.342,P ＜ 0.01) were risk factors of ccRCC.Expression of the Keap1 gene in the patients of the tumor group was significantly lower than that in the patients of the control group (P ＜ 0.05).But there was no obvious difference in expression of the Keap1 gene among samples with different genotypes (P ＜ 0.05).Conclusion:The Keap1 gene rs1048289 could be significantly correlated with ccRCC,and could hopefullv become an effective molecular target for early diagnosis and gene therapy of the patients with ccRCC.