Objective To explore the effects of
down regulation of
osteopontin(OPN)on the bio- logical
behavior of MKN28 and SGC7901
cell lines.
Methods OPN
siRNA was designed according to the relevant
literature and was transfected into the two
cell lines.Fluorescent labeling was used to test the transfected
efficiency.The
down-regulation of OPN
protein was measured by
Western blot.Real-
time PCR was used to test the ratio and
time difference of
down-regulation of OPN
mRNA after
siRNA transfection.The
biological changes before and after OPN
siRNA transfected into these two
cell lines were tested by
flow cytometry(to test
cell cycle and
apoptosis)and MTT
method(to test the prolifera- tion for the consecutive seven days)and the difference between OPN
siRNA transfected or non-transfect- ed
cells was compared using mixed model.The capability of moving and invasion of
cancer cells were tested by Transwell
method and analyzed by t-test.Results The transfected
efficiency of OPN
siRNA were more than 90% in the two
cell lines.OPN
mRNA down-regulated to 47% at the 72th hour in SGC7901,while 40% at the 48th hour in MKN28.The expression of OPN
protein was both down- regulated after
siRNA transfection in the two
cell lines.The proliferation decreased after transfected with OPN
siRNA both in MKN28 andSGC7901(P