The Inhibition And Mechanism Of Genistein Derivatives On Proliferation,Migration,and Invasion Of Huhan Breast Cancer Cell Line Cancer Cells

Objective: To study the effect of genistein derivative WH-20 on the proliferation, migration and invasion of human MCF-7 breast cancer cells, and discuss its preliminary mechanism reasearch,which can provide new ideas for anti-breast cancer drug research.Mothods:1.Cultivating human MCF-7 breast cancer cells in vitro, which was treated with different concentration of WH-20 in different time by MTT method to detect the cell proliferation inhibition rate.After interference with Notch blocking agent(DAPT)?ERa blocking agent(MPP) and ER? blocking agent(PHTPP),We detect the cell proliferation inhibition rate.Plate clone detect the WH-20 on cell's forming ability.The cell cycle effects was detected using WH-20 treated by Flow cytometry.2. human MCF-7 breast cancer cells was treated with different concentration of WH-20.We observe the influence of WH-20 on the cell's migration and invasion by the methods of matrix adhesive experiments, wound healing test and transwell assay. After interference with Notch blocking agent(DAPT)?ERa blocking agent(MPP) and ER?blocking agent(PHTPP),the effects of migration and invasion were evaluated by above expriments,which treated with WH-20.3. Western blot was to detect in different concentration of WH-20 on the MCF-7 cell of the protein expression of cell cycle protein Cyclin D1,related of migration and invasion protein MMP9 and VEGF,estrogen recptor(ERa and ER?),and Notch signaling associated protein such as Notch1,Jagged1,NF-?Bp65,I?Ba. After interference with Notch blocking agent(DAPT)?ERa blocking agent(MPP) and ER? blocking agent(PHTPP)respectively,the above protein was treated by WH-20 used western blot assasy.Results:1. With the increase of WH-20 concentration and time, the inhibitory effect of WH-20 on human MCF-7 breast cancer cells was gradually increased, which could prevent the cell cycle from G1 phase to S phase. and can also inhibit the protein expression of Cyclin D1.2. WH-20 can inhibit the matrix adhesion, migration and invasion of MCF-7 breast cancer cell, and reduce the expression protein of MMP9 and VEGF.3. WH-20 can reduce the expression protein of Notch signaling pathway related Notch1, Jagged1, NF-kBp65, and I?Ba. Adding Notch blocking agent(DAPT) can increase the inhibiton of WH-20 on MCF-7 breast cancer cell proliferation, matrix adhesion, migration and invasion, and the inhibiton of the expression protein of Notch1, Jagged1, NF kappa bp65, I?Ba, Cyclin D1, MMP9 and VEGF were also strengthened.4. The Autodock software simulated successfully the capacity of WH-20 binding with ERa and ER?, and can also increase the expression protein of ERa and ER? by WH-20. Adding ERa blocking agent(MPP) can increase the inhibiton of WH-20 on MCF-7 breast cancer cell proliferation, matrix adhesion, migration and invasion, and the inhibiton of the expression protein of ERa, Notch1, Jagged1, NF kappa bp65, I?Ba, Cyclin D1, MMP9 and VEGF were also strengthened. While adding the ER? blocking agent(PHTPP)was opposite to the effect of the Adding ERa blocking agent(MPP),which can increase the WH-20 on MCF-7 breast cancer cell proliferation, matrix adhesion, migration and invasion,and also incease the expression protein of ER?, Notch1, Jagged1, NF kappa bp65, I?Ba, Cyclin D1, MMP9 and VEGFConclusion: WH-20 can inhibit human MCF-7 cell proliferation, migration and invasion, the major molecular mechanism is through the activation of ERb receptor and inhibition of Notch signaling pathway related protein expression, thereby inhibiting the tumor cell growth related expression protein of Cyclin D1, the migration and invasion of MMP9 and VEGF expression.WH-20 bind the ER? inhibits MCF-7 tumor cell proliferation, migration and invasion by regulation the Notch signaling...