Establishment of integrin #beta#1 knockdown endothelial cell line and study on its proliferation ability

Objective To construct fluorescent small interference RNA(siRNA) plasmid targeting human integrin β1,and to establish integrin β1 knockdown EA.hy926 cell line to investigate the influence of integrin β1 on the proliferation of EA.hy926 cells.Methods Two potential target sequences in integrin β1 open reading frame were selected for the synthesis of DNA templates in vitro.Two DNA strands were annealed into dsDNA,and then the dsDNA was cloned into pGenesil-1 vector.EA.hy926 cells were transfected with recombinant vector and vacant vector.Then the expression of integrin β1 was observed by Western blot.Cell counting was used to analyze the growth curve of cells before and after transfection.Results Restriction endonuclease digestion and DNA sequencing showed that siRNA plasmids based on pGenesil-1 vector were constructed successfully.Western blot revealed that the integrin β1 protein was obviously suppressed by pGenesil 1-integrin β1 transfection.The proliferation of pGenesil 1-integrin β1 transfected EA.hy926 cells was slowed down compared with that of normal ERA.hy926 cells.Conclusion pGenesil 1-integrinβ1 is constructed and integrin β1 knockdown endothelial cell line was established successfully.Downregulation of integrin β1 suppressed the proliferation of human vascular endothelial cells.