| Objective: To explore the effect of Yanghe Tang’s serum containing serum on human breast cancer cell line MDA-MB-231 and its mechanism.To explore the effect of Yanghe decoction on PI3 K / AKT / mTOR signaling pathway,For the clinical use of Yang and soup to provide a theoretical basis for diagnosis and treatment of breast cancer.Methods: Preparation of Yanghe Decoction(Rehmannia 30 g,cinnamon 3 g,ephedra 2 g,antler 9 g,white mustard 6 g,ginger 2 g,raw licorice 3 g,fritillaria 15 g),40 SD The rats were randomly divided into four groups: normal serum control group and Yanghe Decoction medium,high dose group.The normal serum control group was given the same volume of saline for gavage,respectively,the preparation of low,middle and high concentration of drug-containing serum;and use the same way The serum of the control group was prepared.The serum levels of MDA-MB-231 cells were observed by CCK-8 method.The effects of serum on the growth of MDA-MB-231 cells at 24 h,48h and 72 h were observed.The effect of serum containing MDA-MB-231 on the cell cycle distribution and apoptosis was observed by immunocytochemistry.The expression of total PI3 K,p-PI3 K,total AKT,p-AKT,PTEN,total mTOR,p-mTOR and other protein in the cells after 48 h of MDA-MB-231 cells were detected by Western blot.Results: The growth inhibition rate of MDA-MB-231 cells was 0.8%,8.19%and 12.04%,respectively.The inhibitory rate of cell growth was 11.05%,27.28% and51.75%,respectively.The inhibitory rate of cell growth was 2.95%,42.28% and75.49%,respectively.(P <0.05).The positive and negative effects of Yanghe Decoction group were significantly lower than those in the control group(P <0.05),and the difference was significant(P <0.05)Compared with the control group,the absorbance was significantly decreased,with significant statistical difference(p<0.01).Compared with the control group,the percentage of G0 / G1 phase was74.51%,67.93%,60.43% and 52.12% respectively.The proportion of S phase was14.12% in the different dose groups after 48 h treatment with 10%,13.53%,16.06%and 14.94% respectively.The proportion of G2 / M phase was 11.36%,18.53%,23.51% and 32.97% respectively.The three cycles of Yanghe Tang group were compared with the control group It was found that the proportion of G2 / M phase in the three groups was higher than that in the control group(p <0.05 or P <0.01).The results suggest that the effect of yangtang decoction serum on cell cycle distribution is mainly to block cells in G2 / M phase.The percentage of apoptotic cells in each group was significantly higher than that in the control group(P <0.05).The results showed that the percentage of apoptotic cells in the control group was significantly higher than that in the control group The apoptotic rates were 5.97%,7.68% and 9.60%,respectively,which were significantly higher than those in the control group(P <0.01).The expression of PI3 K,Akt and m TOR protein in the cells was significantly higher than that in the control group(P <0.05).The results showed that the levels of PI3 K,Akt and mTOR in the serum were significantly higher than those in the control group(P <0.05).(P <0.05).The expression of PTEN protein increased with the increase of the concentration,and the difference was statistically significant(P <0.05).It was shown that serum at the level of protein was in a concentration-dependent manner to inhibit the phosphorylation of PI3 K,Akt and mTOR proteins and promote the expression of PTEN protein at the protein level.Conclusion: The serum of Yanghe Tang rats can inhibit the proliferation of human breast cancer MDA-MB-231 cells,which can promote the apoptosis of MDA-MB-231 cells,and block the cell cycle in G2 / M phase P-PI3 K,p-Akt and p-mTOR protein expression,and promote the expression of PTEN protein.The above results have a certain time-dose-dependent.This indicates that yangtang decoction has a certain effect on the role of anti-breast cancer,the mechanism may be related to inhibition of PI3 K / Akt / mTOR signal pathway related protein phosphorylation level,thus blocking the cell cycle,speed up cell apoptosis,and ultimately play the anti The effect of the tumor. |
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