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Effects of RNAi-mediated survivin gene silencing prostate cancer cell lines PC3
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Abstract:
To observe the effects of RNAi-mediated survivin gene on silencing prostate cancer cell lines PC3. Methods: Three target gene fragments were cloned into pSilencerTM3. 1-H1 neo vector separately. Three recombinant eukaryotic expression vectors, pSilencer 3. 1-SVV1, pSilencer 3.1-SVV2 and pSilencer 3. 1-SVV3 were successfully constructed, then the recombinant vectors were transfected into prostate cancer cells PC3. After PC3 cells were transfected with recombinant vectors, the interference effects were detected by RT-PCR and Western blot. The apoptosis index and cell cycle of PC3 cells were detected by Flow cytometry. The survival curve of PC3 cells treated with Platinol and the proliferation of PC3 cells were detected by MTT. Results: The results of RT-PCR and Western blot indicated that pSilencer 3. 1-SVV2 and pSilencer 3. 1-SVV3 vectors could knock down the transcription and expression of survivin gene. After PC3 cells were transfected with pSilencer 3. 1-SVV2 and pSilencer 3. 1-SVV3 vectors, the apoptosis index of PC3 cells increased by about 10% -15% , the proliferation of PC3 cells become slowly and the cells number decreased about 30% compared with control groups. The cell number during G1 phases increased 10% and during G2 and S phases the cells number decreased about 5 % . After PC3 cells treated with Platinol, the survival rate of PC3 cells transfected with pSilencer 3.1-SW2 and pSilencer 3. 1-SVV3 vectors decreased by about 35% -45% and apoptosis index increased by about 10% -14% contrast with control groups. Conclusion: The study provides basis for study the function of survivin gene, indicating the survivin may be a new target of gene therapy on prostate cancer.
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