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targeted siRNA against adherence and invasion of human colon cancer cells
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Abstract:
Objective:To construct a small interfering RNA (siRNA) expression vector (psiRNAVEGFR3) targeting vascular endothelial growth factor receptor 3 (VEGFR3) and to investigate the effects of VEGFR3 siRNA on the adherence and invasion of human colon cancer cells. Methods: A siRNA expression vector (psiRNAVEGFR3) targeting VEGFR3were constructed and was used to transfect LoVo cells via lipofectamine 2000. The mRNA and protein expression of VEGFR3were examined after transfection by reverse transcriptase polymerase chain reaction (RTPCR) and Western blotting, respectively. The tumor adhesion ability was detected by cellmatrix adhesion experiment and the invasion ability of tumor cells was evaluated by millicell chamber model. Results: The VEGFR3 siRNA expression vector was successfully constructed. The expression of VEGFR3mRNA and protein was inhibited after psiRNAVEGFR3 transfection. Seventytwo hours after psiRNAVEGFR3 transfection, Western blotting assay showed that the expression of VEGFR3 protein was decreased from (1.26±0.19) to (0.39±0.12)(P＜0.05), the adhesion ability of LoVo cells was also significantly decreased compared with the untransfected group and negative control group (0.407±0029 vs 0.626±0.047,0.621±0.068, P<0.01). The invasion assay demonstrated that the number of LoVo cells penetrating the membrane in the transfection group was significantly lower than those in the untransfected and negative control group (6.38±3.25 vs 24.82±3.44, 23.58±3.73, P<0.05). Conclusion: The siRNA of VEGFR3gene can effectively inhibit the mRNA and protein expression of VEGFR3 in LoVo cells, therefore restraining the adhesion and invasion ability of LoVo cells.
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