南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (01): 11-15.

• • 上一篇    下一篇

siRNA介导的MA PK p42沉默诱导HeLa细胞凋亡

黄辰;刘利英;宋土生;倪磊;宋丽萍;司履生;   

  1. 西安交通大学环境与疾病相关基因教育部重点实验室生物医学基础研究中心医学院遗传学与分子生物学系； 西安交通大学生命科学与技术学院癌症研究所 生命科学与技术学院癌症研究所 陕西 西安 710061； 陕西 西安 710061；
  • 出版日期:2006-01-20 发布日期:2006-01-20
  • 基金资助:
    陕西省自然科学基金(2003C215)

Small interfering RNA-mediated MAPK p42 silencing induces apoptosis of HeLa cells

HUANG Chen, LIU Li-ying, SONG Tu-sheng, NI Lei, SONG Li-ping, SI Lu-sheng Key Laboratory of Environment and Genes Related to Diseases, Cancer Institute, School of Life Science & Technology, Xi’an Jiaotong University, Xi’an 710061, China   

  1. 西安交通大学环境与疾病相关基因教育部重点实验室生物医学基础研究中心医学院遗传学与分子生物学系； 西安交通大学生命科学与技术学院癌症研究所 生命科学与技术学院癌症研究所 陕西 西安 710061； 陕西 西安 710061；
  • Online:2006-01-20 Published:2006-01-20

PDF

1493

摘要： 目的应用siRNA沉默HeLa细胞MAPKP42,研究其对细胞存活的影响.方法体外合成靶向MAPK p42的siRNA-1和siRNA-2,并用脂质体转染HeLa细胞,Western印迹法和免疫组织化学法检测p42MAPK的表达;电镜观察,TUNEL法测定细胞凋亡,Annexin-PI法测定不同凋亡时期细胞的分布.结果siRNA-1和siRNA-2均可使HeLa细胞MAPKp42沉默,使p42MAPK表达下调,与阴性对照组相比分别降低2.5倍和3.2倍.电镜观察证实,siRNA-1和siRNA-2转染组部分细胞丧失表面微绒毛,细胞内空泡增加,细胞核染色质边集.处理组HeLa细胞早期凋亡率显著增加,其中siRNA-1组晚期凋亡率也明显增加.结论siRNA介导的MAPKp42沉默可以诱导HeLa细胞凋亡.

Abstract: Objective To observe the effect of small interfering RNA (siRNA)-induced MA PK p42 silencing on the survival of HeLa cells. Methods Two siRNAs targeting at the MA PK p42 gene and one random siRNA were synthesized respectively by Silencer?siRNA Construction Kit and transfected into HeLa cells by Lipofectamin?2000. The expression of p42MAPK in the transfected HeLa cells was analyzed by Western blotting and immunohistochemistry, and the morphology of cells were observed with electron microscope. TUNEL assay and Annsxin V/PI staining were employed for detecting the cell apoptosis. Results The expression of p42MAPK in the HeLa cells was remarkably suppressed after transfection with the two siRNAs, reduced by about 2.5 and 3.2 folds respectively in comparison with the negative control. Chromatin margination in the cell nuclei were observed in the transfected cells, and TUNEL assay and Annexin V/PI staining further confirmed the occurrence of cell apoptosis. Conclusion In vitro MAPK p42 siRNA-1 and siRNA-2 transfection can specifically silence the gene expression and induce apoptosis of HeLa cells.

中图分类号: