门肺高压症患者骨成形蛋白Ⅱ型受体基因的表达

Expression of bone morphogenetic protein receptor 2 gene in patients with portopulmonary hypertension

目的研究门肺高压症(PPHTN)患者骨成形蛋白Ⅱ型受体(BMPR2)基因的表达。方法确定BMPR2基因13个外显子有效编码序列,使用Premier Primer 5.0和Oligo 6软件设计每个外显子的正、反向测序引物。对4例PPHTN患者、5例门静脉高压症(PHT)对照患者和5例正常对照者的外周血中BMPR2基因的外显子进行PCR扩增和直接测序。结果 1例正常人BMPR2基因的第9号外显子第1 149 bp发生G→A的杂合变异,使第383个密码子由编码甲硫氨酸的ATG变成编码异亮氨酸的ATA(M383I),但BMPR2蛋白质的功能似乎未受影响。此位点不属于目前已知的人类BMPR2基因342个单核苷酸多态性位点之一。其余13例检测结果均正常。结论 PPHTN患者BMPR2基因的外显子编码序列没有发现异常,BMPR2基因的第9号外显子第1 149 bp可能是汉族人的一个新的编码区单核苷酸多态性位点。

Objective To study the expression of bone morphogenetic protein receptor 2（BMPR2） gene in patients with portopulmonary hypertension（PPHTN）.Methods Encoding sequences of all 13 exons in BMPR2 gene were obtained from Genebank of NCBI.Forward and reverse primers of each exon were designed by Premier Primer 5.0 and Oligo 6 software.DNA was isolated from whole blood of 4 PPHTN patients,5 PHT patients and 5 normal volunteers.Protein coding sequences from exon 1 to 13 were amplified from genomic DNAusing the forward and reverse primers.All 13 exons of BMPR2 gene were sequenced directly.Results A heterogeneous missense G→A mutation was detected on 1 149 bp at exon 9 of BMPR2 gene in 1 volunteer,and caused codon 383 change from ATG to ATA（M383I）,but the function of protein unaffected.This mutation could not be categorized as any of the 342 known single nucleotide polymorphisms.There was no other mutation found in the other 13 cases.Conclusion No mutation on encoding sequences of all 13 exons in BMPR2 gene is found in 4 PPHTN patients.The 1 149 bp at exon 9 of BMPR2 gene may be a new location of coding region single nucleotide polymorphisms in Chinese.