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siRNA on expression of angiogenesis-related molecules in gastric cancer AGS cells
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Abstract:
Objective: To study the effect of small interfering RNA (siRNA) targeting Rac1b on the expression of angiogenesis-related molecules in gastric cancer AGS cells. Methods: siRNA targeting Rac1b ( Rac1b siRNA) was synthesized and transfected into AGS cells by lipofectamineTM reagent. The effect of Rac1b siRNA on mRNA and protein expressions of Rac1b in AGS cells was examined by RT-PCR and Western blotting analysis. ELISA and Western blotting analysis were used to assess VEGF production in the supernatants of transfected AGS cells and the expressions of angiogenesis-related molecules such as P53, VHL and HIF-1α in AGS cells under hypoxia situation. Results: DNA sequencing analysis confirmed that the sequence of Rac1b siRNA was correct. Transfection of Rac1b siRNA in AGS cells resulted in a specific decrease in Rac1b mRNA and protein expressions, while failed to knock down the expression of its homologous molecular Rac1. The secretion of VEGF in the supernatant of AGS cells transfected with Rac1b siRNA was significantly inhibited, and this inhibition effect was more obvious under hypoxia condition. Meanwhile, Rac1b siRNA down-regulated HIF-1α protein expression and up-regulated the expressions of angiogenesis-related molecules such as P53 and VHL. Conclusion: Rac1b siRNA can inhibit the mRNA and protein expressions of Rac1b in gastric cancer AGS cells; it can also inhibit VEGF production through regulating the expression of angiogenesis-related molecules such as P53, VHL and HIF-1α under hypoxia condition.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 30770823), and the Natural Science Foundation of Shaanxi Province (No. 2007C222)
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