中文摘要：

目的：构建及鉴定WWOX基因慢病毒载体，观察WWOX基因在人卵巢癌PEO1细胞株中的表达，为进一步研究WWOX基因的作用机制及其功能奠定基础。方法：采用PCR技术从克隆质粒模板中获得全长WWOX基因，将WWOX基因亚克隆到慢病毒载体PCDH—CMV—MCS-EF1-copGFP中，构建慢病毒载体表达质粒PCDH—WWOX，通过双酶切验证后，慢病毒表达质粒与慢病毒包装质粒pPACKH1-GAG，pPACKH1-REV，Pvsv-G共转染293T细胞，获得携带WWOX基因及EGFP基因的重组慢病毒Lenti WWOX，并转染靶细胞人卵巢癌PEO1细胞株。通过RT—PCR和Western Blot验证Lenti WWOX在PEO1细胞株中的表达。结果：测序结果显示成功构建重组慢病毒载体表达质粒PCDH—WWOX；表达质粒与辅助包装质粒共转染包装细胞293T后能产生慢病毒颗粒并能有效感染靶细胞PEO1，转导效率约为60％，有WWOX基因mRNA和蛋白水平的高表达。结论：本实验成功构建了携带WWOX基因慢病毒表达载体，包装成病毒后能有效感染卵巢癌PEO1细胞。

英文摘要：

Objective：To construct the lentiviral vector carrying WWOX gene and investigate their expression in ovarian cancer PEO1 cells. Methods：WWOX gene was acquired from cDNA clone plasmids by RT- PCR, and then subcloned into the expression plasmid of lentiviral vector PCDH-CMV-MCS-EFI-copGFP to generate the lentiviral vector PCDH-WWOX. The lentiviral vector PCDH-WWOX was identified by restriction enzyme. Recombinant lentiviruse was produced by 293T cells following co-transfection of PCDH- WWOX with the packaging plasmids pPACKH1-GAG,pPACKH1-REV and Pvsv-G. The resulting recombinant lentiviruse Lenti WWOX was then used to infect ovarian cancer PEO1 cells. WWOX expression in PEO1 cells was detected by RT-PCR and Western blotting analysis. Results： （1） The recombinant lentiviruse PCDH-WWOX carried the WWOX gene,and it could be expressed in human cell line. （2）The recombinant lentiviruses which carrying WWOX could be product by eo-transfection PCDH-WWOX with packaging plasmid to 293T. （3） The recombinant lentiviruse Lenti WWOX could efficiently infect and deliver WWOX and GFP genes to PEO1 cells. The infection efficiency is about 60% And WWOX gene can be expressed efficiently in PE01 by RT-PCR and Western blotting analysis. Conclusion. The recombinant lentiviruse PCDH-WWOX was produced successfully and it can deliver target gene WWOX to PEO1 cells.