Objective   To establish calcium

calmodulin-associated serine/threonine kinase (CASK) down-regulation ECV-304 cell line and to study the effects of CASK down-regulation on the proliferation of ECV-304 cells.     Methods   The recombinant pGenesil-1-hCASK plasmids containing siRNA targeting CASK gene were transfected into ECV-304 cells. The stable cell strain was screened by adding G418 into cell culture medium. The protein expression levels of CASK gene were detected by Western Blotting. Flow cytometry and MTT method were used to investigate the effects of CASK down-regul

ation on the proliferation of the transfected cells.     Results   CASK down-regulation cell strain

siCASK cell line

was successfully screened. These cells could maintain over 90% green fluorescent protein expression. The decreased expression of CASK could lower but enhance the cell population at G

0

/G

1

phase but G

2

M phase and lead to left-shift of the cell growth curve.     Conclusion   CASK down-regulation cell strain was successfully constructed and CASK down-regulation could enhance the cell proliferation.