| Breast cancer is one of the most common malignant tumors in women due to the high mortality rate of breast cancer metastasis.Formononetin(FMN)is a kind of isoflavonoids derived from the root of herbs,such as the Radix Astragali,Trifolium pratense Linn,Glycyrrhiza Uralensis Fisch,Puerariae Lobata Radix,Spatholobi caulis.It has pharmacological effects including antiinflammation,anti-tumor,cardioprotective activity and neuroprotective activity.In recent years,it has shown that has great potential in inhibiting tumor metastasis.In our study,we investigate that the effects of Formononetin on Proliferation,migration,adhesion,invasion,and related proteins of human breast cancer cell line MCF-7,and according to the mechanism of suppressing tumor metastasis to provide a better development of the antitumor effects of formononetin.Method:The experiment was divided into 5 groups:Negative Control Group(RPMI1640),positive control group(epirubicin hydrochloride,final concentration 0.50 mol/l),low,middle and high dose groups(final concentration 18.75 mol/l,37.50 mol/l,75.00 mol/l,respectively).(1)The antiproliferative effect of formononetin on MCF-7 cells was tested by the CCK-8 assay.(2)Scarification assay was used to detect the effect of formononetin migrational ability in MCF-7 cells.(3)Adhesion assay was used to detect the effect of formononetin adhesion ability in MCF-7 cells.(4)Invasion assay was used to detect the effect of formononetin invasion ability in MCF-7 cells.(5)Western blot was used to analyze the effects of formononetin on the expression of TGF-β1、Ecadherin、Vimentin、MMP-2、MMP-9、ERK1/2、p-ERK1/2、p38and p-p38.Results:(1)Formononetin could suppress the proliferation of MCF-7 cells.The proliferation inhibition rate increased with the formononetin dose increasing,and the value of IC50 was 75 μM for 48 h.The concentration of formononetin in the low,medium and high dose group was 18.75,37.50 and 75.00 μM,respectively.0.50 μM epirubicin was set as the positive control groups.(2)The mobility of each group was detected by cell scratch assay as follows:Blank control group(63.987±0.019)%,Positive control group(29.490±0.004)%,Low dose group(56.537±0.008)%,Medium dose group(29.253±0.004)%and High dose group(14.880±0.001)%.The mobility rates reduced with the formononetin concentration increasing.It was considered that the mobility of MCF-7 cells was suppressed by formononetin.(3)Adhesion rates of each group was detected by cell adhesion experiment as follows:Positive control group(60.25 1±0.054)%,Low dose group(99.320±0.130)%,Medium dose group(59.370±0.118)%and High dose group(34.443±0.038)%.The adhesion rates reduced with formononetin concentration increasing.It was considered that the adhesion of MCF-7 cells suppressed by formononetin.(4)The invasion rates of each group was detected by cell invasion assay as follows:Positive control group(21.250±0.007)%,Low dose group(47.458±0.006)%,Medium dose group(35.156±0.017)%and High dose group(14.881±0.001)%.The invasion rates reduced with the formononetin concentration increasing.It was considered that the adhesion of MCF-7 cells was suppressed by formononetin.(5)Western blot results showed that the expression of E-cadherin was increased by the theophylline.They observed that the inhibitory effects of Vimentin,TGF-1,MMP-2,MMP-9,p-ERK1/2,p-p38 on migration,adhesion and invasion of human breast cancer MCF-7.Conclusion:Formononetin can significantly inhibit the proliferation,migration,adhesion and invasion of human breast cancer MCF-7 cells.The mechanism may be that the expression of Ecadherin,Vimentin and p-ERK1/2,p-p38-related proteins of MMP-2,MMP-9 and Mapk in TGFsignaling pathway are down-regulated by Mantanin,synergistic inhibition of Tumor Migration,adhesion and invasion. |
