Downregulation Of The Expression Level Of CK2 Mediated The Apoptosis Inducation Of The DOdFMG In MCF-7 Cells In Vitro

OBJECTIVE To investigate the effect and mechanism of the Genistein derivative5,4'- Di-n-octoxyl-7-gem-difluoromethylene-genistein(DOdFMG) on the growth and apoptosis of human breast cancer(MCF-7) cells line in vitro, for provide a laboratory and theoretic evidence on searching for a new candidate for tumor chemotherapy .METHODS Soft ager-colon assay was used to test colon formation inhibitory effect of MCF-7 by DOdFMG. PI stain flow cytometry(FCM) was used to analyse the apoptosis after being treated with DOdFMG . Western blotting assay was used to detect the effect of DOdFMG on the CK2,NF-κB ,bcl-2 and Bax protein expression level of MCF-7 cells .RESULTS Soft ager-colon assay showed that the colon inhibitory rate was 21.99% ,41.68% ,63.68% and 48.59% after MCF-7 cells treated with various concentration 3.0, 10.0, 30.0μmol/L DOdFMG and 30.0μmol/L GEN for 6-8d respectively. The DOdFMG caused colon forma-Tion inhibition of MCF-7 cells in a dose-dependent. PI staining flow cytometry showed DOdFMG could induce apoptosis significantly . The activity of inducing the apoptosis of DOddFMG 30μmol/L set is stronger than GEN 30μmol/L set . Western-blotting analysis indicated that after exposures to DOdFMG3.0,10.0,30.0μmol/L for 24h , the protein expressions of CK2 were down-regulated by 12.50%,41.50%,67.30%,the protein expressions of NF-κB were down-regulated by 20.50%,51.47%,71.93%,the protein expressions of bcl-2 were down-regulated by 25.03%,39.27%,58.50% and the protein expression of Bax was up-regulated by 17.60%,22.73%,51.70% respectively in comparison with the control group. After MCF-7 cells were treated with DOdFMG at 30.0μmol/L for 6,12 and 24h, the protein expressions of CK2 were down-regulated by 27.73%,44.8%,65.2%,the protein expressions of NF-κB were down-regulated by 20.5%,48.83%,69.93% ,the protein expressions of bcl-2 were down-regulated by 38.17%,52.30%,71.17% and the protein expression of Bax was up-regulated by 20.50%,65.27%,78.73%, respectively in comparison with the control group. These showed that the expressions of CK2,NF-κB and bcl-2 were decreased and the expression of Bax was increased in a time and dose-dependent manner after treatment with various concentrations of DOdFMG. In comparison with the lead compound Gen, DOdFMG was more effective.CONCLUSION DOdFMG possess a significant inhibitory effect on the cell proliferation and growth of human breast carcinoma cell(MCF-7) in vitro DOdFMG significantly induced apoptosis of human breast cancer (MCF-7 ) cell line. The inhibition of growth and induction of apoptosis of MCF-7 cells by DOdFMG may be associated with downregulation of CK2,NF-κB and bcl-2 protein expression and upregulations of Bax protein expression.