Piwil2基因表达诱导NIH3T3细胞恶性转化  

Malignant transformation of HIN3T3 cells induced by expression of Piwil2 genes

作　　者：李跃波[1] 冯定庆 凌斌[1,2] 程敏[1] 

机构地区：[1]安徽医科大学附属省立医院妇产科,合肥230001 [2]安徽医科大学附属省立医院分子医学实验室,合肥230001

出　　处：《安徽医科大学学报》2013年第5期467-471,共5页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:11040606M176);国家自然科学基金(编号:30901606)

摘　　要：目的通过建立稳定表达外源Piwil2基因的NIH3T3细胞,初步探讨Piwil2基因对NIH3T3细胞生物学特性的调控作用。方法通过脂质体介导的方法和G418的筛选,建立稳定表达Piwil2基因的NIH3T3细胞株,之后利用细胞集落形成实验、细胞增殖实验、细胞周期分析和裸鼠成瘤实验,观察Piwil2基因表达对NIH3T3细胞的生物学特性的影响。结果建立了稳定过表达Piwil2基因的NIH3T3细胞株。与对照组空白载体细胞系相比,过表达Piwil2基因的NIH3T3细胞增殖和集落形成能力增强;细胞周期G0/G1期减少、S期明显升高;接种裸鼠后形成的肿瘤体积显著大于对照组。结论 Piwil2基因在NIH3T3细胞中稳定表达具有诱导正常NIH3T3细胞发生恶性转化的重要生物功能。Objective To establish a mouse fibroblastic cell line stably transfected with Piwil2 gene, and use such cell line to investigate tumor development and progression imposed by the ectopic expression of Piwil2 gene. Methods Eukaryotic expression vector pIRES2-EGFP-Piwil2 was transfected into mouse fibroblast cell line NIH3T3 by lipofectamine. Stable transfect-ants were selected by G418. The integration and expression of Piwil2 gene were analyzed by PCR. And then cell proliferation, cycle and apoptosis in experimental analysis, colony formation and tumor growth experiment were used to observe the impact of Piwil2 gene expression of NIH3T3 on biological characteristics. Results NIH3T3 cell line with stably expression of Piwil2 gene was successfully established and confirmed. Compared with control group, the NIH3T3 cell line with high expression of Piwil2 gene growed quickly and had high capability of colony formation. The S phase and M phase increased significantly. Tumor formation in nude mice was significantly greater than in the control group. Conclusion Expression of Piwil2 gene in NIH3T3 cells can induce malignant transformation of mouse fibroblastic cells both in vitro and in vivo.

关 键 词：Piwil2基因 基因表达 3T3细胞 

分 类 号：R73-3[医药卫生—肿瘤]