Effect of transfected Twist gene on expression of VEGF in human gastric carcinoma cells

Objective To study the effect of transfected Twist gene on expression of VEGF in human gastric carcinoma cells.Methods The re-constructed plasmid PCDNA3.1-Twist was proved by restriction endonuclease technology.MKN28 cells,a human gastric carcinoma cell line,were transfected with PCDNA3.1-Twist plasmid using lipofectanine transfecting technique.Transfected cells were selected with G418.Expression of Twist protein was detected by Western blot in cells transfected Twist gene.VEGF protein of cell culture supernatants was dectected by ElISA methed.The expression of VEGF mRNA was deteced by RT-PCR method.Results MKN28 cells transfected with PCDNA3.1-Twist plasmid showed stronger intracellular expression of Twist protein than MKN28 cells transfected with PCDNA3.1 and MKN28 cells without the transfection.The secrection of VEGF protein in cell culture supernatants of MKN28 cells transfected with PCDNA3.1-Twist plasmid was signifient increased compared with control(P0.05).The VEGF mRNA expression of MKN28 cells transfected with PCDNA3.1-Twist plasmid was higher compared with cells transfected with PCDNA3.1 plasmid and MKN28 cells without transfection.They increased 28.3% and 26.5% respectively.Conclusion Twist can increase expression of VEGF in MKN28 cells.