应用基因测序方法检测哮喘儿童的PTEN基因突变

摘要

Objective To analyze the correlation between phophatase and tensin homolog deleted on chromosone 10 rn(PTEN) and children asthma, to study the relationship of PTEN gene with the number of peripheral blood eosinophils rn(EOS) ,which may explore further pathology and provide some theoretical basis for new therapy of asthma. Methods rnThirty children with asthma(clinical paracmasis) were set as asthma group,thirty healthy children were set as control rngroup. 2ml peripheral vein blood was drawn from every child with empty stomach in early morning. Genomic DNA was rnextracted from each blood sample employing saturated alcohol phenyl and chloroform after preparing smear. Detecting rnthe mutations of exon 5 and exon 8 of PTEN gene by DNA sequencing,meanwhile, counting the peripheral blood EOS.rnMetrology data were analyzed employing χ2 test, enumeration data were analyzed employing t test, the results were any rnlyzed statistical by SPSS 13.0 ,the differences were significant if P<0.05. Results There were mutations at intron 8 ofrnPTEN gene in 83. 3% (25/30)asthma children, there was no mutations in control children. The count of peripheral rnblood EOS in asthma children was obviously higher than that in control children,P<0. 001. The count of peripheral rnblood EOS of asthma children with PETN mutations was higher than that of asthma children without mutations,P< rn0.02. Conclusion There were intron-8 mutations of PTEN gene in most asthma children ,which correlated with the rncount of peripheral blood EOS. PTEN gene may play some roles on the asthma pathology of asthma by refraining the rnactivation and chemotaxis.%目的 分析10号染色体上缺失的磷酸酶与张力蛋白同源基因(phophatase and tensin homolog deleted on chromosone 10,PTEN)突变与儿童哮喘的相关性,及其与外周血嗜酸性粒细胞(eosinophils,EOS)的关系,进一步探讨儿童哮喘的发病机制,并为其新治疗方法的研发提供一定理论依据.方法 设定30例哮喘患儿(临床缓解期)为哮喘组,30例健康儿童为对照组.均于清晨空腹抽取外周静脉血,制备血涂片后提取全基因组DNA.对其PTEN基因第5、第8外显子(exon 5,exon 8)进行基因测序分析,同时进行外周血EOS计数.计量资料采用χ2 test,计数资料采用t检验进行统计分析.结果 83.3%(25/30)哮喘组儿童有PTEN基因第8内含子(intron 8)突变,对照组无基因突变.外周血EOS计数在哮喘组明显高于对照组,差异显著(P＜0.05);发生基因突变的哮喘患儿外周血EOS计数高于无突变的哮喘患儿,差异显著(P＜0.05).结论 哮喘儿童绝大部分存在PTEN基因第8内含子突变,其与EOS的数量相关,PTEN基因可能通过抑制EOS的活化和趋化在哮喘发病中发挥作用.