子痫前期患者胎盘组织PHLDA2基因印迹初步研究  

Placental PHLDA2 Gene Imprinting in Patients with Pre-eclampsia

作　　者：黄桂琼[1] 胡雅毅[1] 王晓东[1] 

机构地区：[1]四川大学华西第二医院妇产科出生缺陷与相关妇儿疾病教育部重点实验室(四川大学),成都610041

出　　处：《四川大学学报（医学版）》2015年第1期104-107,128,共5页Journal of Sichuan University(Medical Sciences)

基　　金：四川省科技厅科技支撑计划项目(No.2011SZ0151)资助

摘　　要：目的观察子痫前期患者胎盘组织母源性印迹基因PHLDA2印迹状态。方法收集21例正常妊娠和19例子痫前期患者胎盘组织,抽提样本DNA,PCR扩增,选择PHLDA2基因组变异比例较高的单核苷酸多态性(SNP)位点:rs13390为外显子1中C/T多态性(PHLDA2-1),rs1056819为外显子2中G/A多态性(PHLDA2-2),直接测序筛选杂合子。将杂合子样本的RNA逆转录合成cDNA,PCR扩增直接测序判断PHLDA2基因印迹状态。结果正常妊娠与子痫前期胎盘PHLDA2-1(外显子1)SNP位点(C/T)均未发现杂合子,全为T/T纯合子;胎盘PHLDA2-2(外显子2)SNP位点(G/A)在子痫前期有4例(4/19)、正常妊娠有5例(5/21)杂合子表达,两组杂合子表达差异无统计学意义(P>0.05)。PHLDA2-2杂合子胎盘组织PHLDA2cDNA均只表达G单等位基因。结论子痫前期胎盘PHLDA2(PHLDA2-1及PHLDA2-2)基因多态性与正常妊娠相似,未发现PHLDA2基因印迹丢失现象。Objective To observe the imprinting status of maternally expressed gene pleckstrin homology-like domain, family A, member 2 （PHLDA2） in placental tissues from patients with pre-eclampsia. Methods Samples of placental tissues were collected from women with normal pregnancy （n= 21） and pre-eclampsia （n= 19）. We examined two single nucleotide polymorphism （SNPs） which are prone to variation in PHLDA2： the C/T polymorphism in exon 1 and the G/A polymorphism in exon 2, corresponding to rs13390 （PHLDA2-1） and rs1056819 （PHLDA2-2）, respectively. DNA PCR-direct sequencing and cDNA RT-PCR-direct sequencing were applied to detect the special-allelic imprinting status of PHLDA2. Results No heterozygote was found in placental tissues in relation to C/T polymorphism in PHLDA2 exon 1. Differences in heterozygote in relation to G/A polymorphism in PHLDA2 exon 2 were found betweenpre-eclampsia （4/19） and normal pregnancy （ 5/21 ）, but without statistical significance. PHLDA2 cDNA from heterozygotes （PHLDA2-2） were all exclusively monoallelically expressed. Conclusion Similargene polymorphism of PHLDA2 （PHLDA2-1 and PHLDA2-2） in placental tissues was found between pre-eclampsiaand normal pregnancies. No loss of imprinting （LOI） of PHLDA2 was found in this study.

关 键 词：基因印迹 杂合子 印迹状态 子痫前期胎盘 PHLDA2 

分 类 号：R714.244[医药卫生—妇产科学]