YANG Hua-an,GOU Xin,ZHENG Chuan-dong,WU Yue,HE Wei-yang.Effects of RNAi-silenced survivin gene on bladder cancer cell lines EJ[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(13):1260-1263.
YANG Hua-an,GOU Xin,ZHENG Chuan-dong,WU Yue,HE Wei-yang.Effects of RNAi-silenced survivin gene on bladder cancer cell lines EJ[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(13):1260-1263.DOI:
Objective To construct eukaryotic expression vectors by using the pSilencer2.0 vector for inhibiting human survivin gene by RNA interference
and to detect the effect of the silenced survivin gene on the growth and apoptosis of EJ cells. Methods Two target gene segments were synthesized and cloned into the pSilencer2.0 vector respectively to construct two recombinant eukaryotic expression vectors: pSilencer2.0-SVV1 and pSilencer2.0-SVV2
both of which were identified by enzyme digestion analysis and DNA sequencing. Then the EJ cells were transfected with the recombinant vectors and the interfering effects were detected by RT-PCR and Western blot. The proliferation of the EJ cells was detected by MTT method and their apoptosis index detected by flow cytometry. Results The results of Enzyme digestion analysis and DNA sequencing showed that two target segments were cloned into pSilencer2.0 vectors. The results of RT-PCR and Western blot indicated that pSilencer2.0-SVV1 and pSilencer2.0-SVV2 vectors could knock down the transcription and expression of survivin gene,but the effect of pSilencer2.0-SVV2 vectors was better than that of the pSilencer2.0-SVV1 vectors. Obvious decrease in growth of the EJ cells was observed after transfected with pSilencer2.0-SVV1 and pSilencer2.0-SVV2 vectors
along with 17% increase of the apoptosis index in the EJ cells transfected with pSilencer2.0-SVV2 vectors. Conclusion The transcription and expression of survivin gene were inhibited effectively in the bladder cancer cell line EJ. After transfected with pSilencer2.0-SVV2 vectors
the apoptosis index of the EJ cells was increased and the growth inhibited.