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Objective:To investigate the effect of RNA interfering on VEGF-C in MDA-MB-231 cells. Methods:Three small interfering RNAs (siRNAa,siRNAb,siRNAc) were prepared. The most efficient one was screened and short hairpin (shRNA) was designed,the recombinant plasmid pGenesil-1/VEGF-C was constructed,and transfected into MDA-MB-231 cells by Lipofectamine TM 2000. RT-PCR,Western-blot an immunohistochemical methods were performed to detect the expression of VEGF-C. Results:RT-PCR results showed that siRNAa,siRNAb,siRNAc could inhibit the growth of MDA-MB-231 cells,among which,siRNAa was the most significant,with an inhibition rate of 72.1%. The recombinant plasmid pGenesil-1/VEGF-C was successfully constructed using shRNA and pGenesil-1. VEGF-C expression was significantly inhibited as determined by RT-PCR,immunocytochemistry staining and Western blot (P<0.05). Conclusion:shRNA RNAi technology could silence the expression of VEGF-C in MDA-MB-231 cells,which suggested that the technology may be one of the effective methods for inhibiting lymphangiogenesis in breast cancer.
Objective: To investigate the effect of RNA interfering on VEGF-C in MDA-MB-231 cells. Methods: Three small interfering RNAs (siRNAa, siRNAb, siRNAc) were prepared. The most efficient one was screened and short hairpin designed, the recombinant plasmid pGenesil-1 / VEGF-C was constructed and transfected into MDA-MB-231 cells by Lipofectamine ™ 2000. RT-PCR, Western- blot an immunohistochemical methods were performed to detect the expression of VEGF-C. Results: RT-PCR results showed that siRNAa, siRNAb could inhibit the growth of MDA-MB-231 cells, among which siRNAa was the most significant, with an inhibition rate of 72.1%. The recombinant plasmid pGenesil-1 / VEGF -C was successfully constructed using shRNA and pGenesil-1. VEGF-C expression was identified as determined by RT-PCR, immunocytochemistry staining and Western blot (P <0.05). Conclusion: shRNA RNAi technology could silence the expression of VEGF- C in MDA-MB-231 cells, which suggested that the technology may be o ne of the effective methods for inhibiting lymphangiogenesis in breast cancer.