摘要
目的探讨采用RNA干扰（RNA interference，RNAi）抑制血管生成素-2（angiopoietin-2，ANG2）基因的表达对人子宫内膜癌细胞Ishikawa细胞的影响。方法采用脂质体Lipofectamine^TM2000介导的方法将针对ANG-2基因的短发卡状RNA（short hairpin RNA，shRNA）表达载体转染到人子宫内膜癌Ishikawa细胞中，逆转录聚合酶链反应及Western印迹法检测ANG-2rnRNA及蛋白的表达；噻唑蓝比色法检测Ishikawa细胞的增殖；荧光显微镜观察细胞形态；流式细胞仪检测细胞周期及凋亡；侵袭实验检测其对侵袭能力的影响。结果人子宫内膜癌Ishikawa细胞ANG-2mRNA及其蛋白质表达水平均显著降低；增殖被抑制，抑制率63．11％；细胞凋亡增加，细胞凋亡率与空白对照组和阴性对照组相比明显增高；细胞生长减慢，阻滞于G1期的增多，S期细胞减少；侵袭能力明显下降。结论靶向ANG-2的shRNA能成功下调ANG-2基因的表达，抑制人子宫内膜癌Ishikawa细胞生长和侵袭。
Objective To investigate the effect of RNA interference mediated angiopoietin-2（ ANG- 2 ）gene silencing on human endometrial carcinoma cell line ]shikawa. Methods Short hairpin RNA （shRNA） targeting ANG-2 gene was designed and transfected into Ishikawa cells with LipofectamineTM 2000. The mRNA and protein expression level of ANG-2, proliferation, morphological changes, apoptosis, cell cycle and invasive ability of the cells after transfection were analyzed. Results The shRNA targeting the human ANG-2 gene effectively decreased the expression orANG-2 on both mRNA and protein level, the proliferation inhibition rate of the Ishikawa cells was 63.11%, cell apoptosis was induced, and the cell cycle was arrested in G1 phase. The apoptotic rate of the Ishikawa cells in the transfected group was significantly higher, and the invasive ability was decreased markedly, than that of control groups. Conclusion The shRNA targeting human ANG-2 gene could reduce ANG-2 expression, inhibit cellular growth and invasion in Ishikawa cells in vitro.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2009年第3期249-253,共5页
Chinese Journal of Medical Genetics
基金
江苏省社会发展计划研究与发展资助项目（BS2004007）
