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  • Title: Construction of survivin siRNA expression vector and its regulation on cell cycle and proliferation in MCF-7 cells.
    Author: Li LP, Liang NC, Luo CQ.
    Journal: Ai Zheng; 2004 Jul; 23(7):742-8. PubMed ID: 15248905.
    Abstract:
    BACKGROUND & OBJECTIVE: Survivin is specifically overexpressed in tumor tissues. Many reports have shown that the abrogation of its functions is useful for tumor therapy. RNA interference is a new technique that proved to be effective for suppressing gene expression. The aim of this study was to construct a siRNA expression plasmid against gene survivin, and then to assess its functions on breast cancer cells MCF-7. METHODS: A survivin siRNA plasmid was constructed using a mU6pro vector contained U6 promotor, the expression change of survivin was examined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis after the plasmid had been transfected into MCF-7 cells, and the effect of the plasmid on the cell cycle and cell proliferation was analyzed by flow cytometry and MTT method,respectively. RESULTS: Survivin siRNA plasmid knocked down survivin expression in MCF-7 cells obviously, arrested the cell cycle in G1 phase, inhibited the cell proliferation significantly, and promoted cell apoptosis in a tendency. CONCLUSION: Survivin expression was knocked down at mRNA and protein levels by RNAi.
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