Extraction Of Panax Ginseng Protein And Its Anti-Tumor Effects On Breast Cancer Cells MCF-7

Objective:In order to select the best extraction method of Panax ginseng protein and compare the effects of different growth years on ginseng protein content,the anti-tumor effect and mechanism of Ginseng Protein on human breast cancer MCF-7 cells were studied.Methods:Water-soluble protein of Panax ginseng was extracted by acetone precipitation,PEG6000 precipitation,PEG20000 precipitation,phenol extraction and Trizol extraction,and the proteins of 3,4 and 5-year-old ginseng were compared.The protein content was determined by Bradford method and molecular weight was identified by SDS-PAGE electrophoresis.Different concentrations of ginseng protein were applied to human breast cancer MCF-7 cells,CCK-8 method was used to determine the proliferation activity on cells,the changes of cell cycle were detected by PI staining and flow cytometry,the apoptotic morphology of the cells was observed by normal light microscopy and DAPI staining,Annexin V/FITC double staining method was used to further detect cell apoptosis,QPCR and Western Blot experiments were used to detect the effects of gene and protein levels,cell migration was detected by cell scratch healing test and Transwell chemotaxis chamber test,and the mechanism of ginseng protein was studied.Results:The ginseng protein extracted by acetone precipitation has the highest purity,and the electrophoresis band is the clearest and complete.By comparing the protein content of ginseng in 3,4 and 5 years,it was found that the protein content of 3-year-old ginseng was the lowest,and that of 5-year-old ginseng was the highest.The results of the activity study indicate that Ginseng protein can significantly inhibit the proliferation of MCF-7 cells,The IC₅₀ values of 24,48 and 72 hours were 5.21,2.59 and 1.11 mg/ml respectively;Ginseng protein can block the cell cycle of MCF-7 at G0/G1 phase,and ginseng protein can cause apoptosis in MCF-7 cells and induce apoptosis,its mechanism of action is to induce the apoptosis of MCF-7 cells by affecting the expression of Bax and Bcl-2 at the gene and protein levels,increasing the ratio of Bax/Bcl-2,increase the permeability of the mitochondrial membrane,thereby inducing apoptosis of MCF-7 cells;Ginseng protein can effectively inhibit the migration of MCF-7 cells.Conclusion:Acetone precipitation method is a better method for protein extraction of ginseng,and the protein content of ginseng increases with the growth year;Ginseng protein has significant anti-tumor effect on human breast cancer MCF-74 cells,the mechanism of action is to inhibit cell proliferation and inhibit tumor cell migration by regulating the cell cycle of MCF-7;Simultaneously inhibit tumor cell migration and growth;It can also promote the apoptosis of cancer cells,its mechanism is that the expression of Bax and Bcl-2genes is affected by mitochondrial pathway,which mediates the programmed apoptosis of cancer cells;At the same time,it inhibits the migration and growth of cancer cells.This experiment provides a reliable research basis for the future development and application of ginseng protein in the fields of food,health care products and pharmaceuticals.