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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: Total RNA was extracted from human normal ovarian tissue and the human WWOX gene was amplified by reverse transcriptase polymerase chain reaction (PCR). The eukaryotic expression vector was constructed and transfected into human ovarian cancer cell line A2780. RESULTS: The human WWOX gene was successfully cloned and showed complete agreement with the sequence provided by Genbank (NM-016373). WWOX mRNA expression in cancer cells was significantly increased after transfection with eukaryotic expression vector pCMV-WWOX in human ovarian cancer cell line A2780 (P <0.01). Conclusion: Human WWOX gene was successfully cloned from human normal ovarian tissue, which laid the foundation for further study on the role of WWOX in the development and progression of ovarian cancer and its targeted gene therapy.