Comparison of inhibitory effects of microRNA and siRNA on survivin gene expression and apoptosis in prostate cancer
AIM:To design and construct microRNA expression rectors, and to compare microRNA and siRNA inhibitive effects on survivin gene expression. METHODS : Based on survivin. gene sequence, targeted pre-microRNA oligonucleotides were designed and synthesized respectively, and cloned into established microRNA expression vector pPRIME. PC-3 cells were transiently trans-fected with correct constructs of microRNA and siRNA and the inhibitory effects were determined by PT-PCR and Western blot. The apoptosis index of PC-3 cells was detected by flow cytometry and electron microscope. RESULTS: Double enzyme digestion a-nalysis and DNA sequencing results confirmed that the sequence of survivin specific microRNA expression vector was correct. Survivin mRNA and protein expression level in PC-3 cells were specifically suppressed after transfection with recombinant plasmids. The results of flow cytometry indicated that the apoptosis index increased obviously in pPRIME-SV and pSilencer3.1-SVV2 recombinant plasmids. The results of cellular structure and cell organ in electron microscope displayed corresponding changes. The inhibi-tive effects of microRNA expression vector were better than those of siRNA expression vector. CONCLUSION: Both microRNA and siRNA expression vectors inhibit gene expression and induce apoptosis of prostate cells, but microRNA expression vector a-chieves better effect than siRNA expression vector.
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万方数据
维普
