人WWOX基因的克隆及其真核表达载体的构建

Cloning of human WWOX gene and construction of its eukaryotic expression vector

目的:克隆人WWOX基因并构建其真核表达载体。方法:从人正常卵巢组织中提取总RNA,经逆转录聚合酶链反应扩增人WWOX基因,并构建其真核表达载体,经测序鉴定后转染人卵巢癌细胞系A2780。结果:成功克隆了人WWOX基因,与Genbank提供的序列(NM-016373)对比显示完全一致。真核表达载体pCMV-WWOX转染人卵巢癌细胞系A2780后,癌细胞的WWOXmRNA的表达水平显著增高(P<0.01)。结论:从人正常卵巢组织中成功克隆了人WWOX基因,为进一步研究WWOX在卵巢癌发生和进展中的作用及其靶向基因治疗奠定了实验基础。

Objective：To clone human WWOX gene and construct its eukaryotic expression vector.Methods：The total WWOX RNA was abstracted from human normal ovarian tissues,then human WWOX gene was amplified by RT-PCR,its eukaryotic expression vector was constructed,human ovarian cancer cell line A2780 was transfected after gene sequencing.Results：The human WWOX gene was successfully cloned and it was identical to the reported WWOX sequence（NM-016373） provided by Genbank.After the eukaryotic expression vector pCMV-WWOX was transfected into ovarian cancer cell line A2780,the expression level of WWOX mRNA increased significantly（P0.01）.Conclusion：The human WWOX gene is successfully cloned from human normal ovarian tissues,which provides a experimental basis for investigating the effect of WWOX in the occurrence and development of ovarian cancer and targeted gene therapy.