千金子对表达人G250基因小鼠renca细胞增殖的影响

发布时间：2018-09-07 20:30

【摘要】：目的观察中药千金子对表达人G250基因小鼠renca细胞增殖的影响。方法利用DNA重组技术将成功扩增的人G250基因编码区序列定向插入到pRNAT-U6.1/Neo质粒真核表达载体中,得到重组质粒pRNAT-U6.1/Neo-G250,将该质粒转染renca小鼠细胞,经G418筛选获得抗性单克隆,扩大培养后经Western印迹鉴定人G250基因的表达。将该细胞在不同浓度含千金子二萜醇(111μg/ml、333μg/ml、1 000μg/ml、3 000μg/ml、9 000μg/ml)的培养基中培养,MTT法观察千金子二萜醇对表达人G250基因小鼠renca细胞体外增殖情况的影响。结果与空白对照组相比,24 h后实验组细胞增殖速度减慢,并随浓度增加增殖速度减慢越明显,空白组与3 000μg/ml及9 000μg/ml浓度组比较差异有明显统计学(P=0.006,P=0.000)。结论中药千金子使表达人G250基因小鼠renca细胞增殖能力降低,renca细胞体外生长被显著抑制。
[Abstract]:Objective to observe the effect of Qianjinzi on the proliferation of renca cells expressing human G250 gene in mice. Methods the successfully amplified coding region of human G250 gene was inserted into the eukaryotic expression vector of pRNAT-U6.1/Neo plasmid by DNA recombination technique. The recombinant plasmid pRNAT-U6.1/Neo-G250, was transfected into renca mouse cells, and the resistant monoclonal was obtained by G418 screening. The expression of human G250 gene was identified by Western blotting after expanded culture. The cells were cultured in different concentrations of 1 000 渭 g / ml 1 000 渭 g/ml to observe the effect of 1 000 渭 g / ml 1 000 渭 g/ml on the proliferation of renca cells expressing human G250 gene in vitro. Results compared with the control group, the proliferation rate of the experimental group decreased after 24 h, and decreased more obviously with the increase of the concentration. There was a significant difference between the blank group and the 3 000 渭 g/ml and 9 000 渭 g/ml groups (P0. 006, P0. 000). Conclusion Qianjinzi can significantly inhibit the proliferation of renca cells expressing human G250 gene in vitro.
【作者单位】：河南省中医院河南中医药大学第二附属医院泌尿外科;南方医科大学附属南方医院泌尿外科;兰州军区总医院泌尿外科;
【基金】：河南省科技攻关重点项目(142102310287)
【分类号】：R285

【参考文献】