Induction Of The Growth Inhibition And Apoptosis In Human Breast Cancer Cell Line By 5, 4-Di-n-octoxyl-7-gem-difluoromethylene-genistein In Vitro

Purpose: To investigate the effect of the Genistein derivative, 5,4'-Di-n-octoxyl-7-gem-difluoromethylenegenistein(DOdFMG) on the growth and apoptosis of human breast cancer(MCF-7) cell line in vitro.Methods: MCF-7 cells line and HBL-100 cells line were treated with various concentration of DOdFMG.. MTT assay was used to examinate the influence of growth in MCF-7 cells and HBL-100 cells. Cell counting method afer trypan blue staining was used to test the influence of growth curve in MCF-7 cells and HBL-100 cells.The influence of colone formation in MCF-7 cell were evaluated by plate clone method. Apoptosis of MCF-7cells was determined by AO/EB double fluorescence staining and flow cytometry after PI staining, and cell cycle was analyzed using flow cytometry.Results: MTT assay indicated that DOdFMG significantly suppressed proliferation of MCF-7 cells in dose-dependant and time-dependant manner, which were obviously stronger antitumor activity than GEN. The IC₅₀ of MCF-7 cellsat 24 hour,48 hour,72 hour were 20.05μmol/L,15.23μmol/L,11.14μmol/L respectively. DOdFMG has a weaker inhibitory action of HBL-100 cells, the IC50 of HBL-100 cells at 24 hour,48 hour,72 hour were 78.05μmol/L,69.71mol/L,65.54μmol/L respectively.Its selectivity is stronger than GEN ,having smaller toxicity to the normal cells.The SI of MCF-7 cells at 24 hour,48 hour,72 hour were 3.89,4.58,5.88 respectively. These results are consistent with the results of Cell counting method after trypan blue staining plotting growth curve in MCF-7 cells and HBL-100 cells. Plate clone formation method manifested that DOdFMG obviously repressed multiplication of MCF-7 cell colonies in concentration-dependant manner, compared with GEN has a conspicuously stronger antitumor activity. Flow cytometry after PI staining and AO/EB double fluorescence stain data showed that the apoptosis of MCF-7cells induced and cell cycle was arrested at S stage and G₂ stage by DOdFMG in a concentration-dependent manner. In comparison with GEN 100μM group, activity of inducing the apoptosis of DOddFMG 40μM group is stronger.Conclusion: (1)DOddFMG can significantly inhibit MCF-7 cells growth ,having little effect in HBL-100 cells and it is a new chemical entities with more relative selectivity for treat human breast cancer. (2) DOddFMG can induce apoptosis ,arrest cell cycle in G₂ and S phase of MCF-7 cells .