脑胶质瘤病人遗传易感基因单核苷酸多态性的研究

Study on the single nucleotide polymorphysims of the susceptibility genes in patients with glioma

目的筛查与脑胶质瘤发病相关基因的单核苷酸多态性,进一步寻找胶质瘤易感基因。方法收集入院脑胶质瘤病人的血液。应用PCR扩增的方法对肿瘤相关的基因的特定片段进行扩增。利用变性高效液相色谱技术进行筛查。双向DNA直接测序,在GENBANK进行对比找到突变的碱基,并进行分析。结果变性高效液相色谱筛查后表明,在HLA-DMA基因中检测到了已知杂和性突变,并发现新的突变,HLA-DMA4EC杂合性缺失突变。结论变性高效液相色谱技术在筛查基因的SNPs方面显示了其快速、高通量和准确的特点。HLA-DMA基因第4号外显子第111位碱基缺失突变是一个新的SNP,可能是胶质母细胞瘤的易感突变之一。

Objective To screen the SNPs of the relative genes responsible for glioma pathogenesis. Subsequently to find the susceptibility gene for brain glioma. Methods The samples of blood and tumor tissues were collected from all of the patients and controls, genomic DNA was extracted from whole blood, and then special fragments of tumor relative genes, such as HLA-DMA, were amplified with PCR. We screened these fragments with the technique of dHPLC, and directly sequencing was performed with the fragments which have mutations. In order to find oat the mutation basic, the sequence results were compared with the data in GenBank. Results After the scree,fing with the method of dItPLC, Mutations were found in the fragments, wltieh belong to HLA-DMA. Among them, only the mutation of LOH of HLA-DMA4E111C has statistic significance compared with the classification of the gliomna and the lifestyle of the patients. This mutation is related to glioblastoma and AⅢ, and also to contacting with pesticide. Conclusion dHPLC is a good method for scree,fing SNPs of gene with the excellence of speediness and high flux. The mutation in exon 4C111 of HLA-DMA is a novel variation found in this study.