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Objective: eukaryotic translation initiation factor 4E (EIF4E) as cell growth and differentiation important regulatory factors, and tumor occurrence is closely related. In this study, gene transfection of human EIF4E small interfering RNA (small interfering RNA) expression plasmid, explore its breast cancer MDA-MB-231 cells biological behavior. Methods: The synthetic gene targeting EIF4E shRNA, and cloning of eukaryotic expression vector pGPU6/GFP/Neo to construct pGPU6/GFP/Neo-EIF4E recombinant plasmid by restriction enzyme digestion and sequencing analysis of the use of liposomes LipofectamineTM2000 Identification of the correct recombinant plasmid transfected into human breast cancer MDA-MB-231 cells, using RT-PCR, Western blotting, immunocytochemistry, immunofluorescence assay before and after transfection EIF4E, CyclinD1, VEGF-C, MMP-2, MMP- 9 mRNA and protein expression; MTT and colony formation assay to detect cell proliferation and colony formation rate; Hochest33258 staining cells; flow cytometry cell cycle; Transwell experiment, cell scratch assay cell invasion and migration changes. Results: restriction analysis and sequencing confirmed eukaryotic recombinant plasmid pGPU6/GFP/Neo-EIF4E, and stably transfected human breast cancer MDA-MB-231 cells. RT-PCR, Western blotting, immunofluorescence, immunocytochemistry showed that recombinant plasmid group MDA-MB-231 cells EIF4E, Cyclin D1, VEGF-C, MMP-2, MMP-9 mRNA and protein expression were significantly decreased (P lt; 0.05). Recombinant plasmid slow cell growth, proliferation, reduced ability to significantly reduce the number of colony-forming, with the blank control group and empty vector group, the difference was statistically significant (P lt; 0.05). Hchest33258 staining to the nucleus concentrated, deeply stained, and changes in cell membrane characteristics of apoptosis, apoptotic index was significantly higher (P lt; 0.05). Flow cytometry showed that after transfection increased the percentage of cells in G1 phase (71.3 ± 0.467 vs 53.1 ± 0.431)%, S phase cells was significantly reduced (12.53 ± 0.125vs 26.47 ± 0.376)% (P lt; 0.05). Invasion assay results, the small number of cells under the membrane chamber blank group (341.7 ± 21.47), expression plasmid group was (79.43 ± 11.21), the difference was statistically significant (P lt; 0.05); migration experiments, a small chamber film the number of cells of the control group (639 ± 47.34), expression plasmid group (298.23 ± 20.01), a significant difference between (P lt; 0.05). Experimental results show that cell scratch recombinant plasmid group was significantly decreased cell migration (P lt; 0.05). Conclusion: Recombinant eukaryotic expression plasmid pGPU6/GFP/Neo-EIF4E significantly reduced EIF4E gene in breast cancer MDA-MB-231 cells, to a certain extent, inhibit breast cancer cell proliferation, invasion and migration, promote cell apoptosis, so that more cells remain in the G1 phase, S phase cells decreased, and its mechanism may be associated with reduced VEGF-C, Cyclin D1, MMP-2, MMP-9 expression, suggesting that EIF4E siRNA sequences may become the treatment of an effective means of breast cancer.
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