Study On The Screening And Action Mechanism Of Berberine Anti-tumor Activity

The mechanism that berberine promotes the apoptosis of colon cancer cells was found by comparing with CMap.Berberine has certain anti-tumor activity.By comparing the gene expression profile data of berberine and CMap database,the mechanism of berberine against tumor was predicted.Then CCK-8 was used to determine the tumor cells used to validate the anti-tumor mechanism of berberine.Methods:First,the RNA of MCF-7 Human breast cancer cells was treated with berberine to make the gene chip,wherein the chip type was Affymetrix Human Genome U133A 2.0,and the gene expression profile data of berberine was determined.By berberine gene expression profile data,compared with that of CMap database to predict berberine may be as a kind of Hsp90 inhibitors,may also be the HDAC inhibitor,possibly through the inhibition of PI3K/AKT/mTOR pathway to inhibit the growth of tumor cells.And then through the CCK-8 method determination of berberine of several laboratory commonly used the antitumor activity of tumor cells,and the SW480 colon cancer cells antitumor activity is best,so follow-up experiments to choose SW480 cells.Cell apoptosis,mitochondrial membrane potential and cell cycle were detected by flow cytometry.And through Cellular thermal shift detection in SW480 cells,whether berberine and Hsp90 proteins,by Western blot test whether berberine affect protein Hsp90 customers,part of the HDAC subtypes and PI3K/AKT/mTOR pathway of protein expression.Finally through the CCK-8 method and the calculation of the combination index to analysis whether berberine and 17-AAG?a kind of Hsp90 inhibitors?or SAHA?a selective HDAC inhibitor?combined in SW480 cells have synergy and whether the three combined have synergy.Results:Among the tumor cells detected,berberine had the best anti-tumor activity of the people colon cancer cells of SW480,with the IC₅₀ as 3.436+0.193?M.Berberine can promote SW480 cell apoptosis through mitochondrial apoptosis,and berberine in SW480 cells can inhibit the PI3K/AKT/mTOR pathway,and thus promote the cells in autophagy,block cell cycle at G1 phase of SW480 cells.Berberine is combined with 17-AAG or SAHA in SW480 cells,with a certain synergistic effect.All three are used simultaneously,and there is a certain synergy,and the synergistic effect is more obvious.Otherwise,due to the mentioned in the previous reports in some tumor cells,Notch1 pathway can down-regulate the PTEN protein to affect the PI3K/AKT/mTOR pathway.Therefore,we studied whether berberine could affect Notch1 pathway and the expression of PTEN protein in SW480 cells.Methods:We used Western blot to detect the change of protein expression in Notch1 pathway and the expression of PTEN protein.Results:Berberine can inhibit the expression of Jagged1,Notch1,NICD,and Hes-1 proteins in the Notch1 pathway in SW480 cells,and the expression of the upstream PTEN protein in the PI3K/AKT/mTOR pathway can be up-regulated by inhibiting the expression of Hes-1 protein.Conclutions:1.Berberine in SW480 cells can inhibit the PI3K/AKT/mTOR pathway,and thus promote SW480 cell apoptosis through mitochondrial apoptosis,promote SW480cells in autophagy,block the cell cycle at G1 phase.2.In SW480 cells,the combination of berberine and Hsp90 inhibitors or HDAC inhibitors has synergistic effect.The combination of the three also has synergistic effect,and the synergistic effect is more obvious.3.Berberine can inhibit the Notch1 pathway in SW480 cells,and may therefore up-regulate the expression of PTEN protein to inhibit the PI3K/AKT/mTOR pathway.