The Effect And Related Molecular Mechanism Of Total Flavonoids From Lycium Barbarum L. On The Proliferation,Migration And Invasion Of Breast Cancer Cells

Flavonoids are widel y distributed in plants and traditional Chinese medicines.They have prominent anti-cancer effect on breast cancer,lung cancer,leukemia, gastric cancer and so on.At the functional level,they alter cell proliferation,differentiation,apoptosis,angiogenesis,migration,and invasion.Lycium Barbarum L.is traditional Chinese medicinal materials that have anti-inflammatory and anti-tumor effects.Flavonoids are one kind of its main ingredients.However,the effect of total flavonoids derived from Lycium Barbarum L.?TFLb?on breast cancer has not been reported yet.Objective:To investigate the effects and mechanism of total flavonoids from Lycium barbarum L.?TFLb?on the types of breast cancer cells,include proliferation,migration and invasion.Methods:Two breast cancer cell l ines MCF-7 and MDA-MB-231 were selected,MCF-7 is ER positive and MDA-MB-231 is a trip-negative breast cancer cell line.After treated with different concentrations of TFLb,1)the cell proliferation was detected by CCK-8 assay.Meanwhile,MCF-10A was used as a control.2)The changes of cell migration and invasion were detected by cell Wound Healing assay and Transwell assay.3)Use flow cytometry to detect the cell cycle and apoptosis.4)Use q PCR to detect the change of m RNA expression levels of PCNA and other molecular indicators.5)Western Blot was used to detect the changes of molecular marker proteins like PCNA,MMP-2.We also detect the changes of some key proteins in cell signaling pathway include Rho A,Rac1/Cdc42,ER?NF-?B,?-catenin,Akt and m TOR.to explore the mechanism.Result:1)The inhibitory effect of TFLb on breast cancer in a dose-dependent manner.After treated with 48h,more than 90%proliferation was inhibited when200?g·m L^-1 TFLb to MCF-7 or 100?g·m L^-1 TFLb to MDA-MB-231,both concentrations did not inhibit proliferation of normal mammary epithelial cells MCF-10A.2)Treat MCF-7 with 45,90?g·m L^-1 or MDA-MB-231 with30,60?g·m L^-1 TFLb inhibit cell not proliferation but migration and invasion.3)45,90?g·m L^-1 TFLb arrest MCF-7's cell cycle at G0/G1 phase,30,60?g·m L^-1TFLb arrest MDA-MB-231's cell cycle at G2 phase.4)Treat MCF-7 with135?g·m L^-1 or MDA-MB-231 with 90?g·m L^-1 TFLb promote breast cancer cell apoptosis,and change the ratio of intracellular Bcl-2/Bax protein.5)Akt,beta-catenin and phosphorylation of Akt1 were decreased when treat MCF-7 with45,90,135?g·m L^-1 or MDA-MB-231 with 30,60,90?g·m L^-1 TFLb after 48h.Moreover,the contents of Rho A,Rac1/Cdc42,NF-?B and ER?proteins were decreased in MCF-7 cells.In MDA-MB-231,m TOR and Rac1/Cdc42 were reduced only with 90?g·m L^-1 TFLb.Conclusion:Below the concentrations which are innocuous to normal mammary epithelial cells MCF-10A,TFLb have many similar effects of two kind breast cancer cells.Thes e effects include inhibit proliferation,migration,invasion,arrest cell cycle and promote apoptosis.The effective concentration and action pathway of TFLb on MCF-7 cells and MDA-MB-231 cells are different,yet?-catenin and Akt are common effect sites in both cells.