CAJ | 학술논문

目的探讨采用RNA干扰(RNA interference,RNAi)抑制血管生成素-2(angiopoietin-2,ANC-2)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LipofectamineTM 2000介导的方法将针对ANG-2基因的短发卡状RNA(short hairpin RNA,ahRNA)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-2 mRNA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-2 mRNA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.
목적 탐토채용RNA간우(RNA interference,RNAi)억제혈관생성소-2(angiopoietin-2,ANC-2)기인적표체대인자궁내막암세포Ishikawa세포적영향.방법 채용지질체LipofectamineTM 2000개도적방법장침대ANG-2기인적단발잡상RNA(short hairpin RNA,ahRNA)표체재체전염도인자궁내막암Ishikawa세포중,역전록취합매련반응급Western인적법검측ANG-2 mRNA급단백적표체;새서람비색법검측Ishikawa세포적증식;형광현미경관찰세포형태;류식세포의검측세포주기급조망;침습실험검측기대침습능력적영향.결과 인자궁내막암Ishikawa세포ANG-2 mRNA급기단백질표체수평균현저강저;증식피억제,억제솔63.11%;세포조망증가,세포조망솔여공백대조조화음성대조조상비명현증고;세포생장감만,조체우G1기적증다,S기세포감소;침습능력명현하강.결론 파향ANG-2적shRNA능성공하조ANG-2기인적표체,억제인자궁내막암Ishikawa세포생장화침습.
Objective To investigate the effect of RNA interference mediated angiopoietin-2(ANG-2)gene silencing on human endometrial carcinoma cell line Ishikawa. 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 of ANG-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 ANC-2 gene could reduce ANG-2 expression, inhibit cellular growth and invasion in Ishikawa cells in vitro.