88例胆囊结石患者线粒体tRNA基因突变分析

Analysis of mitochondria tRNA gene mutations in 88 cases with cholecystolithiasis

目的筛选胆囊结石患者线粒体tRNA基因的突变位点,探讨线粒体tRNA与胆囊结石的相关性。方法收集88例胆囊结石患者和376例正常对照者的血液标本,提取外周血基因组DNA,PCR扩增,产物纯化,高通量测序方法测序并分析结果。结果胆囊结石患者在性别、BMI、Glu、SBP、LDL 5个项目上与对照组比较,差异有统计学意义(P<0.05)。胆囊结石组中共发现15个线粒体tRNA基因突变,涉及变异位点38个。tRNA基因保守性>75%的有12个。tRNA基因二级结构分析中,共有9个突变导致WC碱基配对改变,2个产生了新的碱基配对,7个导致碱基配对消失。结论通过对线粒体tRNA基因测序结果进行评估,推测以下8个位点可能与胆囊结石病程相关:tRNAPheG625A、tRNACysA5810G和T5814C、tRNAIleA4295G、RNAAlaT5587C、tRNAGlyA10005G、tRNALeu(CUN)A12280G、tRNAThrA15896G;线粒体tRNA基因是胆囊结石的突变热点区域。

Objective To detect the mutation sites of mitochondria tRNA gene in patients with cholecystolithiasis,and to investigate the correlation between mitochondria tRNA and cholecystolithiasis. Methods Blood samples from 88 patients with gallbladder stones and 376 normal controls were collected; genomic DNA was extracted from peripheral blood for PCR amplification and product purification; high-throughput sequencing was performed and the sequencing results were analyzed.Results Compared with the control group,there was statistical significance on the differences of sex,BMI,Glu,SBP and LDL in the patients with cholecystolithiasis（ P〈0. 05）. 15 mutations of mitochondrial tRNA gene were found in the cholecystolithiasis group,including 38 variants. There were 12 mutations with CI 75%. In the analysis of the secondary structure of the tRNA gene,a total of 9 mutations led to WC base pairing change,2 mutations resulted in new base pairs,and 7 mutations resulted in base pairing loss. Conclusion It is speculated that the following 8 sites may be the mitochondrial tRNA variation sites associated with the course of cholecystolithiasis,including tRNAPheG625 A,tRNACysA5810 G and T5814 C,tRNAIleA4295 G,RNAAlaT5587 C,tRNAGlyA10005 G,tRNALeu（ CUN）A12280 G,tRNAThrA15896 G. Mitochondrial tRNA is a hot spot of cholecystolithiasis mutation.