Blocking effect of vectorª²based siRNA on stathmin gene expression in human HeLa cells

    WANG Yan, WU Bing, SU Haiª²Chuan, LIU Li, LIN Fang, ZHANG Huiª²Zhong

    Department of Clinical Diagnosis, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China

    ¡¾Abstract¡¿ AIM: To explore the blocking effect of siRNA on the expression of stathmin gene in HeLa cell line using siRNA eukaryotic expression vector. METHODS: Two stathmin siRNA cDNAs were synthesized according to the stathmin gene sequence and cloned into the vector pSilencer4.1ª²CMVª²neo and named pSilencerª²S1 and pSilencerª²S2 respectively, which were further identified by restriction endonuclease digestion analysis and DNA sequencing. HeLa cells were then transfected with pSilencerª²S1 and pSilencerª²S2. After G418 selection, the cells were selected and the interfering effect was detected by RTª²PCR. RESULTS: Restriction endonuclease digestion analysis and DNA sequencing results showed that the 2 target segments were cloned into pSilencer4.1ª²CMV neo vector respectively. The results of RTª²PCR indicated that both siRNA vectors could successfully knock down stathmin gene expression. CONCLUSION: The vectorª²based siRNA on stathmin gene can effectively knock down stathmin gene expression, which has the potential of being applied in gene therapy against malignant tumors. ......