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Archive > Volume 17 Issue 1 > 2010,17(1):51-56. DOI:10.3872/j.issn.1007-385X.2010.1.010 Prev Next

Basic Research

RNAi targeting AKT1 and PI3K P85 suppresses proliferation of breast carcinoma MCF-7 cells

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    Abstract:
    Objective:To investigate the effect of RNA interference (RNAi) targeting AKT1 and PI3K P85 on the proliferation and invasion of breast carcinoma MCF7 cells. Methods:The recombinant adenovirus expression vector, which contained short hairpin RNA (shRNA) targeting open reading frames of AKT1 and PI3K P85(rAd5siAKT1siPI3K), was transfected into human breast carcinoma MCF7 cells. AKT1 and PI3K P85 mRNA and protein expressions were detected by realtime PCR and Western blotting analysis. The expressions of PCNA, cyclinD1, and P53 were also detected by Western blotting analysis. The proliferation and apoptosis of MCF7 cells were measured by MTT, flow cytometry and 2dementinal and 3dementional matrigel assay. Results:Recombinant adenovirus vector rAd5siAKT1siPI3K dramatically downregulated AKT1 and PI3K P85 mRNA and protein expressions in MCF7 cells; the downstream factors PCNA and cyclin D1 were also downregulated, while P53 was upregulated. Growth of MCF7 cells was inhibited by over 50% in rAd5siAKT1siPI3K group as measured by MTT assay, and cell cycle was arrested in G1/G0 phase compared with untransfected and rAd5siCtrl transfected groups. Cell growth on matrigel matrix showed normal cell shapes, while the cells in rAd5siAKT1siPI3K transfected group were detached from the matrix or grew in scattered clustering patterns, forming only small aggregates. Conclusion:shRNA targeting AKT1 and PI3K P85 can significantly downregulate the expression of AKT1 and PI3K P85 in breast carcinoma MCF7 cells, and inhibit the growth of MCF7 cells in vitro.

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    Project Supported:
    Project supported by Major State Basic Research Development Program of China (973 Program) (No.2009CB918903), and the National Natural Science Foundation of China (No.30670802), and the Applied Basic and Advanced Research Program of Tianjin (No.09JCZDJC1

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