Axin基因对人肾母细胞瘤细胞生物学特性的影响  

Biological effects of Axin gene on human Wilms' tumor cells

作　　者：陈广生[1] 安鲁[1] 李娟[1] 张强[1] 惠军朋 

机构地区：[1]西安市儿童医院病理科,西安710002

出　　处：《陕西医学杂志》2016年第5期619-622,共4页Shaanxi Medical Journal

基　　金：陕西省自然科学基金资助(2010JM4026)

摘　　要：目的:研究Axin基因对人肾母细胞瘤细胞G401生物学特性的影响为人肾母细胞瘤的分子病理机制及治疗、预后提供基础研究依据。方法:采用基因工程的方法构建含Axin基因的真核表达载体pIRES2-EGFP-Axin,稳定转染G401细胞,建立高表达的G401细胞系。用MTT实验、克隆形成能力实验、侵袭能力实验三种方法检测Axin基因对G401细胞生物学特性的影响。结果:重组载体pIRES2-EGFP-Axin经PCR和双酶切鉴定结果显示:在约2500bp大小处出现一条特异性条带,转染G401细胞后,经Western blotting结果显示:Axin蛋白在G401细胞中成功表达,同时,转染了pIRES2-EGFP-Axin质粒的G401细胞生长速度显著减慢(P<0.05);G401细胞的克隆形成能力和侵袭力均显著降低(P<0.05)。结论:我们成功构建了重组真核表达载体PIRES2–EGRP–Axin,且通过Western blotting检测筛选获得稳定转染的G401细胞系,最后Axin体外实验对G401细胞生物学特性影响表明:Axin基因与肾母细胞瘤的发生发展有一定关系。Objective：Research effects of biological characteristics of Axin gene on human Wilms tumor cell G401 to molecular pathogenesis,treatment and prognosis of Wilms tumor provide basic research basis.Methods：Constructed eukaryotic expression vector pIRES2-EGFP-Axin by genetic engineering methods,transfected stably G401 cell,establishing of G401 cell lines of high expression,By MTT assay,colony forming ability experiments and invasion experiment three methods detect effects of biological characteristics of Axin gene in G401 cell.Results：Recombinant vector pIRES2-EGFP-Axin by PCR and double digestion results showed that about 2500 bp appeared in a specific band,after transfected G401 cell,Western blotting results showed：Axin protein successfully expressed in G401 cell.Meanwhile MTT assay in vitro experiments showed that：transfected pIRES2-EGFP-Axin G401 cell growth slowed significantly（P〈0.05）,Colony formation and invasion experiment results also show：after transfected plasmid pIRES2-EGFP-Axin the G401 cell colony formation and invasion were significantly reduced（P〈0.05）.Conclusion：We successfully constructed the recombinant eukaryotic expression vector PIRES2-EGRP-Axin,and screened stably transfected cell lines through Western blotting analysis,the last Axin in vitro effects on the biological characteristics of G401shows：a certain relationship between Axin gene with the occurrence and development of Wilms＇ tumor.

关 键 词：肾母细胞瘤/治疗 基因/治疗应用 细胞分裂 @Axin @G401 

分 类 号：R737.11[医药卫生—肿瘤]