APOL1基因变异的检测及其与高血压肾病的初步关联研究

Establishment of whole-gene sequencing for APOL1 gene variations and its preliminary interconnection with hypertensive renal disease

目的:建立载脂蛋白L1(APOL1)基因变异的检测方法,初步观察APOL1基因变异与高血压肾病(HRD)的关联性。方法:检索确认APOL1基因序列,设计第2~7外显子的7对引物。提取人外周血样本基因组DNA,行PCR扩增,扩增产物经琼脂糖电泳鉴定、纯化,行全基因测序。检测10例HRD患者APOL1基因,进行非同义突变的功能评价,分析其与HRD的关联性。结果:成功扩增了APOL1基因第2~7外显子,测序证实含目标外显子序列。10例HRD患者中发现APOL1基因变异2例,这些变异可能具有致病性。结论:成功建立了APOL1基因的全基因测序方法。HRD患者存在APOL1基因变异,需积累病例进一步证实两者关联性。

AIM：To establish whole-gene sequencing method for apolipoprotein L1（APOL1）gene,and to examine a potential interconnection between APOL1 variations and hypertensive renal disease（HRD）in Chinese Han population. METHODS：According to confirmed gene sequence,7 pairs of primes for APOL1 gene exons 2 - 7 were designed.Peripheral blood was collected and total genomic DNA was prepared for PCR. PCR products were purified and Sanger sequencing analysis was performed in 10 patients with HRD. Genetic variations were visualized and evaluated by Mutation Surveyor software. The association of APOL1 variations with HRD was evaluated. RESULTS：Six APOL1 gene exons were amplified successfully,and their PCR products were identified to contain target sequences by sequencing. APOL1 gene exons 2 ~ 7 were amplified and variations were detected by Sanger sequencing. Two nonsynonymous mutations of APOL1 gene,c. 382 C 〉 T（p. R128C）and c. 533 C 〉 T（p. A178V）,were found in 2 of 10 HRD patients,and their functional impacts were predicted to be likely pathogenic. CONCLUSION：Whole-gene sequencing analysis showed that variants of APOL1 gene existed in Chinese Han HRD patients. It’s necessary to collect more samples to evaluate potential association between APOL1 gene variations and HRD.