Genistein Down-regulates Notch-1 Expression And Causes Hedgehog Signaling Pathway Inactivation Through SHH And HHIP Induction In Pancreatic Cancer Cells

Objective To investigate the influence of genistein on Notch-1,SHH and HHIP gene expression and study its effect on the proliferation of pancreatic cancer cells.Methods Human pancreatic cancer cell line BXPC-3 was cultured in RPMI-1640 media supplemented with 10% fetal bovine serum in a 5% CO2 atmosphere at 37℃.The BXPC-3 cells were treated with genistein and then extracted the total RNA and protein .Semi-quantitative RT-PCR was used to detect the expression of Notch-1,SHH and HHIP mRNA. Notch-1 and SHH protein was checked by western-blotting. MTT assay was used to detect proliferation of BXPC-3 cells. The cell cycle of BXPC-3 cells was studied by Propidium iodide (PI) method and flow cytometry.Results Notch-1 mRNA and protein expression were inhibited after pancreatic cancer cells were treated by 20ug/ml genistein for 48 hours. Simultaneously, SHH mRNA and protein expression were decreased and HHIP mRNA expression was increased. Genistein can inhibit the proliferation of BXPC-3 cell line and block the cell cycle. The inhibiting rate was 67.17% and 52.77% cells were hindered in G2/M stage after pancreatic cancer cells were treated by 20ug/ml genistein for 48 hours.Conclusions Genistein can down-regulate Notch-1 expression and inactivate Hedgehog signaling pathway, and then inhibit the proliferation of pancreatic cancer cells. The inhibitions of Notch-1 combines with Hedgehog signaling pathway will be a new therapeutic method of pancreatic cancer probably.