Inhibitory effect on human leukemia K562 cells by RNA interference of stathmin gene

Objective:To study the RNA interference-mediated inhibition of stathmin gene on the proliferation of human leukemia K562 cells. Methods: Two target gene segments were synthesized and cloned into vector respectively to construct two recombinant eukaryotic expression vectors: pSilencer4.1-CMV neo-S1 and pSilencer4.1-CMV neo-S2.The two recombinant vectors were identified by enzyme digestion analysis and DNA sequencing. Then K562 cells were transfected with pSilencer4.1-CMV neo-S1and pSilencer4.1-CMV neo-S2 ,then subjected to Neomycin selection. In Neomycin-resistant cells, the interference effect was detected by RT-PCR. The proliferation of transfected K562 cells was examined by MTT assay. Results: Enzyme digestion analysis and DNA sequencing showed that the target segments were cloned into pSilencer4.1-CMV neo vector respectively. The result of RT-PCR indicated that both vectors could knock down the transcription of stathmin gene. At the same time, the growth of transfected cells was decelerated. Conclusion: stathmin gene silencing by RNA interference contributed to a distinctive inhibition of the proliferation of human leukemia K562 cells in vitro.