Journal of Southern Medical University ›› 2006, Vol. 26 ›› Issue (12): 1806-1811.

Previous Articles     Next Articles

RNA interference-mediated inhibition of survivin expression in Hela cell line by siRNA expression vector targeting survivin gene

JIANG Hong-chao1,2,SUN Qiang-ming1,ZHAO Dan2,SUN Mao-sheng1,LU Lin2,LI Hong-jun1 1Department of Molecular Biology,Institute of Medical Biology,Chinese Academy of Medical Sciences and Peking Union Medical College,Kunming 650118,China;2Graduate School of Kunming Medical College,Kunming 650031,China   

  1. 中国医学科学院/中国协和医科大学医学生物学研究所分子生物室; 昆明医学院研究生部; 中国医学科学院/中国协和医科大学医学生物学研究所分子生物室 云南昆明650118昆明医学院研究生部; 云南昆明650031; 云南昆明650118;
  • Online:2006-12-20 Published:2006-12-20

Abstract: Objective To prepare small interfering RNA(siRNA)targeting survivin for inhibition of endogenous survivin gene expression in Hela cell line and evaluate its effect on promoting Hela cell apoptosis.Methods The recombinant plasmid pshRNA-survivin-1 and pshRNA-survivin-2 were constructed and transfected into Hela cells,in which the expression level of survivin was determined by immunofluorescence staining and survivin gene transcription detected by semi-quantitative RT-PCR.Results Introduction of the plasmids pshRNA-survivin-1 and pshRNA-survivin-2 into Hela cells resulted in efficient and specific inhibition of survivin expression as demonstrated by immunofluorescence staining.Semi-quantitative RT-PCR showed that mRNA transcription of survivin gene was reduced.In contrast,the control plasmid did not exhibit any inhibitory effect on the protein expression and mRNA transcription of survivin gene.PI-Annexin V staining indicated an apoptosis rate of the transfected Hela cells of(36.02±2.12)%(P<0.01)and(35.29±2.02)%(P<0.01),respectively.Conclusion The prepared siRNA targeting survivin gene is capable of inducing marked inhibitions of survivin protein expression and RNA transcription and significant enhancement of apoptosis in Hela cells,which shed light on a new strategy in gene silence therapy targeting survivin.

CLC Number: