人脑胶质母细胞瘤中核因子-κB抑制因子α多态性的研究

The clinical analysis of nuclear factor-κB inhibitor-α polymorphism in glioblastoma

目的探讨人脑胶质母细胞瘤中核因子-κB抑制因子d（NFKBIA）蛋白水平及其与NFKBIA mRNA表达水平、变异、拷贝数变异的关系。方法收集24例胶质母细胞瘤组织标本，应用聚合酶链反应（PCR）扩增及直接测序检测NFKBIA变异，Western blot检测NFKBIA蛋白表达，实时定量PCR检测NFKBIA拷贝数变异及mRNA表达。结果胶质母细胞瘤中存在NFKIBA单核苷酸多态性rs1957106（CC／CT／TT）；胶质母细胞瘤中NFKBIA蛋白表达量在CT基因型和TT基因型显著低于CC基因型：CT和TT：（10．1±1．2）％、CC：（40．1±2．8）％；胶质母细胞瘤中NFKBIA蛋白表达量和NFKBIA mRNA表达水平均显著低于非瘤组织：肿瘤组织中NFKBIA的相对表达分别为：（28．2±12．5）％、（0．44±0．19），非瘤组织中NFKBIA的相对表达分别为：（64．3±1．7）％、（0．78±0．12）；而且含有CT基因型和TT基因型的胶质母细胞瘤NFKBIA mRNA表达水平明显低于含有CC基因型的胶质母细胞瘤：CT和TT：0．31±0．11，CC：0．54±0．18；P〈0．01；含有CT基因型或TT基因型的10个胶质母细胞瘤中有7个的拷贝数显著低于含有CC基因型的肿瘤，而这些胶质母细胞瘤同时有低NFKBIA蛋白及低NFKBIA mRNA表达水平。结论NFKBIA单核苷酸多态性rs1957106与NFKRTA低蛋表达、低拷贝数密切相关。

Objective The aim of the present study was to explore nuclear factor-κB inhibitor-α （NFκBIA） protein levels and their relationship with mRNA levels, mutation and copy number variation of NFκBIA in glioblastomas. This will lay a foundation for further clarifying the role of NFKBIA in the development of glioblastomas. Methods We analyzed 24 human glioblastoma samples and 8 samples of noncancerous brain tissue for mutations, copy number variation and expression （protein and mRNA levels） of NFKBIA. We used polymerase chain reaction（PCR） amplification and direct sequencing to detect mutaions of NFKBIA, Western blotting to detect expression of NFκBIA protein, real-time quantitative PCR to detect copy number variation and mRNA expression of NFκBIA. Results We genotyped one single nucleotide polymorphism rs1957106 in NFKBIA in glioblastomas. NFKBIA protein levels detected were significantly lower in glioblastomas harboring the rs1957106 CT and TT genotypes than in samples harboring the rs1957106 CC genotype： CT and TT： （ 10. 1 ± 1.2）%, CC： （40. 1 ±2. 8）%. Moreover, NFKBIA protein levels were significantly lower in glioblastomas than in non-cancerous brain tissues： tumor： （28.2 ± 12. 5） % , non-cancerous brain tissue： （64. 3 ± 1.7） %. NFKBIA mRNA levels were lower in glioblastomas compared with non-cancerous brain tissue, tumor： 0. 44 ±0. 19, non-cancerous brain tissue： 0. 78 ± 0. 12. Furthermore, NFKBIA mRNA levels were significantly lower in glioblastomas harboring the rs1957106 CT and TT genotypes than in samples harboring the rs1957106 CC genotype （CT and TT： 0. 31 ±0. 11, CC： 0. 54 ±0. 18）, P 〈0. 01. The relative copy number of NFKBIA was significantly lower in 7 of the 10 glioblastomas, all of which had the rs1957106 CT and TT genotypes and low NFKBIA protein lev els. Conclusion The minor allele for one synonymous single nucleotide polymorphism in NFKBIA, rs1957106, was associated with lower nrotein exnression and lower cony number of NFKBIA.