| PURPOSE To investigate the effects of the Genistein derivative, 5,4'-Di-n-octoxyl-7-gem-difluoromethylenegenistein (DOdFMG) on inhibition of growth and induction of apoptosis of human ovarian cancer(CoC1) cells line in vitro. METHODS CoC1 cells were cultured in vitro. MTT assay was used to determine the effect of DOdFMG on the proliferation of CoC1 cells. The trypan blue exclusion method was used to investigate the growth inhibitory effect of DOdFMG in CoC1 cells. AO/EB fluorescence staining was used to observe the morphologic changes of apoptosis induced by DOdFMG in CoC1 cells. The protein level and activity of NF-κB,Bcl-2 and Bax were examined by Western blot to explore the molecular mechanism of anti-ovarian cancer action of DOdFMG .RESULTS The MTT assay showed DOdFMG significantly suppressed proliferation of CoC1 cells in a concentration-dependant manner, that the proliferation inhibitory rate was 39.1%,47.7%,59.1%,53.4% respectively after CoC1 cells treated with various concentration3.0,10.0,30.0μM of DOdFMG for 48h, and its IC50 was 11.9μM. DOdFMG at 10.0μM could inhibit the growth of CoC1 cells and prolong CoC1 cell multiple time from 37.8h to 50.6h.Typical morphologic changes of apoptosis could be observed after treatment with DOdFMG by fluorescence microscope using AO/EB fluorescence staining. Western-blotting analysis indicated that after CoC1 cells treated with DOdFMG at 0.3,3.0,30.0μM for 24h the protein level of NF-κB,Bcl-2 was down-regulated 6.83%,21.7%,38.3% ,11.6%,25.4%,46.2% in comparision with the control group ,whereas the protein level of Bax was up-regulated 16.5%,37.6%,82.7%.CONCLUSION (1)5,4'-Di-n-octoxyl-7-gem-difluoromethylenegenistein possess the inhibitory effect of the proliferation and growth of CoC1 cells in a dose-dependent manner . (2)5,4'-Di-n-octoxyl-7-gem-difluoromethylenegenistein can significantly induce apoptosis of CoC1 cells. (3)The inhibition of growth and induction of apoptosis of CoC1 cells by 5,4'-Di-n-octoxyl-7-gem-difluoromethylenegenistein may be associated with downregulation of NF-κB ,Bcl-2 protein level and upregulation of Bax protein expression . |
