摘要
目的了解乙型肝炎病毒(HBV)感染胎盘组织基因表达的改变,确定与 HBV 感染胎盘相关的基因,以探讨 HBV 宫内感染的分子机制。方法选择2002年1月至2003年12月,在西安交通大学第一医院未临产以剖宫产分娩的30例 HBsAg、HBV DNA 双阳性(实验组)和30例 HBsAg、HBV DNA 双阴性(对照组)妇女的胎盘组织,采用抑制消减杂交技术(SSH)建立 HBV 感染胎盘组织cDNA 消减文库,经反向斑点杂交技术证实差异表达基因。RT-PCR 验证部分基因在 HBV 感染胎盘组织中的表达。结果两组差异表达基因35个,对其中含有插入片段的阳性克隆进行测序分析,获得33个已知基因序列,两个未知基因;并初步判定磷脂酰肌醇3激酶(PI3K)基因在 HBV 感染的胎盘组织中表达明显增强。结论 HBV 感染胎盘时,基因表达可发生明显变化;PI3K 基因表达变化,对了解 HBV 宫内感染的分子机制有重要意义。
Objective To screen differentially expressed genes in placentas with hepatitis B virus （HBV） infection and to discuss the molecular mechanism of HBV intrauterine infection. Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group. The suppression subtractive hybridization （SSH） technique was used. Total RNAs of placenta tissue of the study group were mixed as the tester, and total RNAs of placenta tissue of the control group were mixed as the driver. A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems. Amplifications of the library were carried out with E. coli strain DH5α by reverse spot hybridization. RT-PCR confirmed that phosphatidylinositol 3-kinase （PI3K） was up-regulated in placenta tissue with HBV infection. Results Colony PCR showed that the clones contained 200 - 1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics. Thirty three known genes and 2 genes with unknown function were obtained. RT-PCR prehminarily confirmed that PI3K gene was up-regulated in HBV infected placenta. Conclusions The differentially expressed genes in placentas with hepatitis B virus （HBV） infection using SSH technique has been screened out successfully. These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation, and signal conduction-antiapoptosis pathway. This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2007年第2期76-78,共3页
Chinese Journal of Obstetrics and Gynecology
基金
陕西省科技计划(2005K15-G2)
