Rap2b基因表达对NIH3T3细胞集落形成及细胞伤口愈合能力的影响

目的了解Rap2b基因对NIH3T3细胞集落形成及细胞伤口愈合能力的影响,为探讨该基因在人肺癌发生中的作用提供实验依据。方法以人肺CDNA为模板克隆出Rap2b基因片段,构建pcDNA3.1-Rap2b真核表达载体,稳定转染NIH3T3细胞,利用集落形成试验和细胞伤口愈合试验,观察细胞集落形成及细胞伤口愈合能力的改变。结果与转染空质粒的细胞对比,转染pcDNA3.1-Rap2b质粒的细胞集落形成数目明显增加(P0.01),损伤修复愈合增快。结论 Rap2b基因表达可显著增加NIH3T3细胞集落形成,并增快该细胞的损伤愈合速度。提示Rap2b基因表达可能具有促进NIH3T3细胞增殖和恶性转化功能。

Effects of Rap2b gene on foci formation and wound-healing of NIH3T3 cells

Objective To explore the effects on foci formation and wound-healing of NIH3T3 cells,and to provid the experimental evidence of its function.Methods DNA from human lung cells was extracted and amplification of Rap2b gene was done by PCR.Eukaryotic expression vector pcDNA3.1-Rap2b was constructed and was stable transfected into NIH3T3 cell followed with foci formation assay and wound-healing assay.Results The number of the foci formation of NIH3T3 cell transfected by eukaryotic expression vector pcDNA3.1-Rap2b was increased remarkably in foci formation assay ( P < 0.01),and NIH3T3 cells transfected by eukaryotic expression vector pcDNA3.1-Rap2b were quickly heal up in wound-healing assay.Conclusion The extrinsic expression of Rap2b could transform NIH3T3 cell using foci formation assay and wound-healing assay.Rap2b gene might play an oncogenic role in tumorigenesis.