肾细胞癌患者抑癌基因VHL双等位基因失活的测定

Analysis of biallelic inactivation of the von Hippel-Lindau tumor suppressor gene VHL in patients of renal cell carcinoma patient

目的探讨肾细胞癌患者中抑癌基因VHL的双等位基因失活发生情况。方法在41例肾细胞癌患者中提取肿瘤和正常组织DNA。采用单链聚合酶链反应(PCR)和测序法检测肿瘤组织中VHL基因的突变情况。采用PCR限制性片段长度多态性法检测VHL基因内部的2个单核苷酸多态性(SNP)位点,在2个位点的杂合子中通过对比肿瘤组织和正常肾组织分析VHL基因的杂合性丢失(LOH)情况。结果在肾细胞癌中51%(21/41)发生VHL基因突变,42%(8/19)发生VHL基因LOH,基因突变和LOH发生具有显著一致性(r=078),在37%(7/19)的肾细胞癌中发生VHL双等位基因失活。结论肾细胞癌中存在由VHL基因突变和LOH导致的VHL双等位基因失活现象,VHL双等位基因失活发生率为37%。

Objective To investigate biallelic inactivation of the von Hippel-Lindau tumor suppressor gene (VHL) in patient of renal cell carcinoma (RCC) patient. Methods We extracted tumor and normal DNA from 41 RCC patients. Mutation of VHL gene from tumor tissue was detected from tumor tissue by polymerase chain reaction (PCR) and direct sequencing. Two single nucleotide polymorphism (SNP) sites located in VHL gene were analyzed by PCR restriction fragment length polymorphism, and loss of heterozygosity (LOH) was analyzed for VHL gene by comparing between tumor with normal tissue. Results Mutation and LOH of VHL gene was found in 51% (21/41) and 42% (8/19) of RCC patients respectively. LOH was highly associated with mutation positive tumors(r=0.78) and VHL biallelic inactivation was detected in 37% of RCC patients.Conclusion Biallelic inactivation of VHL gene occurs in RCC due to VHL mutation and LOH, and its frequency rate is 37%.