背景与目的:前期研究显示,FXR1基因是从卵巢上皮癌cDNA文库中筛选到的相关抗原基因,FXR1抗原蛋白的自身抗体可用于早期卵巢癌的诊断.本研究的目的是分析上调上皮性卵巢癌相关抗原基因FXR1表达对卵巢癌细胞系HO8910生物学行为的影响.方法:采用半定量RT-PCR法检测在不同卵巢组织中,FXR1 mRNA的表达,并分析其与临床病理的关系.RT-PCR扩增FXR1 CDS,构建FXR1真核表达载体,脂质体介导下转染HO8910细胞,经G418筛选获得稳定转染株,检测转染后细胞相应生物学行为.结果:FXR1 mRNA在卵巢癌组织中的相对表达水平比卵巢良性肿瘤组织及正常卵巢组织均增高(P＜0.05);FIGO分期Ⅰ～Ⅱ期和高分化癌组织中FXR1 mRNA相对表达水平低于Ⅲ期和中-低分化组织(P＜0.05);成功构建了FXR1基因的真核表达载体,并稳定转染了HO8910细胞.生长曲线显示上调FXR1基因表达可促进细胞的生长;克隆形成实验显示上调FXR1基因表达可促进细胞增殖.基质黏附实验显示,上调FXR1基因表达可加强细胞对基质的黏附能力,流式细胞周期检测亦发现与对照(80%)相比,转染FXR1基因后G1期细胞(67.1%)比例明显减少,这说明转染FXR1后细胞的增殖更旺盛.同时发现FXR1对细胞侵袭、迁移的能力无明显影响.结论:上调FXR1基因表达可能有促进卵巢癌细胞的生长、增殖、加强细胞对基质黏附能力的作用.%Background and purpose: Primary study showed that FXR1 is an associated antigen gene screened from epithelial ovarian cancer (EOC) cDNA library. The autoantibody of EXR] antigen can be used to diagnose early stage ovarian cancer. The goal of the study is to investigate the biological behavior change of EOC cells HO8910 by up-regulated EOC associated antigen gene FXR1 expression. Methods: The semi-quantitative RT-PCR was used to detect the FXR1 rnRNA expression in ovarian cancer tissues, benign tissues and normal ovarian tissues. The relationship between The FXR1 mRNA expression and clinical-pathology factor also was analyzed. FXR1 CDS was amplified by RT-PCR. The PCR product was digested with corresponding restriction enzymes and subcloned into pCDNA3.1 plasmid. HO89I0 cells transfected withpCDNA3.1/FXRl or pCDNA3.1 were selected in G4I8 to generate [[()8910/pC:i)NA3.1/[;XR1 and i[()8910/pCi)NA3.1. Ceils growth curve, clone formation assay, adhesion assay, in vitiv migration, invasion assay, and flow cytometry assay were examined. Results: The FXR1 mRNA expression in ovarian cancer was higher than those in benign and normal control (P展开▼
