nm23-H2基因抑制卵巢癌细胞侵袭转移及调节网络初探

Research of function and gene regulation network of invasion and metastasis related nm23-H2 gene of epithelial ovarian cancers

目的研究上皮性卵巢癌转移相关nm23-H2基因功能及其调控网络,为最终揭示卵巢癌侵袭转移分子机制和晚期卵巢癌基因治疗提供理论依据。方法将nm23-H2重组质粒pCDNA3/nm23-H2转染人卵巢癌细胞系SKOV3.ip1,经G418抗性筛选及RT-PCR鉴定得到稳定转染阳性克隆H2-18,设计体外黏附实验及人工基底膜侵袭实验比较转染前后细胞黏附侵袭特性的改变。应用2 747点人类肿瘤相关Oligo芯片检测转染前后细胞基因表达的改变,寻找与nm23-H2相关下游调控肿瘤相关基因,并探讨其调节网络。结果用重组质粒pCDNA3/nm23-H2转染SKOV3.ip1细胞。转染前后细胞在体外的增殖能力没有显著变化,转染后细胞的黏附与侵袭基底膜能力明显降低。经人类肿瘤相关Oligo芯片检测转染前后细胞发现55个二倍差异表达基因,在上调基因中以转录调节、信号转导、分化发育及细胞凋亡类基因为多;下调基因中以信号传导及黏附运动功能类基因为主。结论nm23-H2基因具有抑制卵巢癌侵袭转移的作用,并且这一作用的实现是通过对一系列下游肿瘤相关基因的调控来完成的。基因芯片技术结合普通分子生物学技术是后基因组时代肿瘤相关基因功能研究及探索基因调控网络的有效手段。

Objective To investigate the function and gene regulation network of invasion and metastasis related nm23-H2 gene of epithelial ovarian cancer,with the hope of providing theory basis for the molecular mechanism of ovarian cancer invasion and metastasis,and seeking for a new approach of gene function study in postgenome era.Methods Epithelial ovarian cancer cells SKOV3.ip1 were transfected with human recombinant plasmid pCDNA3/nm23-H2.The stable transfected clone H2-18 with G418 resistance was further confirmed by RT-PCR of the nm23-H2 gene expression.The adhesion and invasion ability to basilar membrane was compared between H2-18 and vector-alone transfected C2.And human cancer Oligo chip with 2 747 genes was used to explore the relevant down-streaming genes and the gene regulation network of nm23-H2.Results The adhesion and invasion ability to basilar membrane of H2-18 was reduced compared with vector-alone transfected clone C2,while the multiplication capacity unchanged.The expression of 55 genes was found changed after nm23-H2 transfection.According to the gene function category,they mainly belong to the categories of transcription regulation,signal transduction,adhesion and motility and differentiation.Conclusions Nm23-H2 gene can be a metastasis inhibitor of epithelial ovarian cancer through regulations of other down-streaming genes.Microarray combined with bioinformatics methods is an effective way for gene function research in postgenome era.