| Objective: The serum containing yanghe decoction(YHD)was prepared to interfere with MCF-7 cells of breast cancer,and then the proliferation of MCF-7 cells was observed.Through the mitogen-activated protein kinase(p38)/signal transduction and transcriptional activator 3(STAT3)signaling pathway,the mechanism of YHD in the treatment of breast cancer was explored,providing experimental basis for subsequent trials and experimental basis for clinical application.Methods: 1.Preparation of(YHD): 1 g·m L-1(YHD)was prepared by boiling the medicinal materials purchased from the yanghe decoction.2.Preparation of serum containing(YHD): 20 female SPF SD rats were randomly divided into a control group(distilled water)and a low-dose,medium-dose and high-dose(YHD)group(6 g·kg-1,12 g·kg-1,24 g·kg-1).After three days of intragastric administration,blood was collected from the abdominal aorta and centrifuged to prepare the drug-containing serum.3.Cell culture: MCF-7 cells were resuscitated,DMEM medium containing fetal bovine serum and dual antibodies was added,and cultured in CO2 incubator at 37?.4.Cell intervention: the logarithmic growth cells were divided into the control group,the low,medium and high dose groups,and the corresponding drug-containing serum was added for intervention.Cell proliferation and cytotoxicity assay(CCK-8)was used to detect the effect of serum containing YHD on MCF-7 cell proliferation.The cell cycle of MCF-7 cells was determined by cell cycle experiment.DAPI staining was used to detect the effect of serum containing YHD on the nuclear morphology of MCF-7 cells.Flow cytometry(Annexin,V-FITC /PI double staining)was used to detect the effect of serum containing yanghetang on apoptosis of MCF-7 cells.Western blot was used to detect the effects of serum containing YHD on the expression of p38,Survivin,b-cell lymphoma/leukaemia-xl(bcl-xl),phosphorylation signaling and transcriptional activfactor 3(p-stat3)and STAT3 in MCF-7 cells.Quantitative real-time PCR was used to detect the effect of serum containing YHD on m RNA expression of p38,STAT3,Survivin and bcl-xl in MCF-7 cells.Results: CCK-8 results showed that,compared with the control group,the serum containing YHD inhibited MCF-7 cell proliferation,which was related to time and dose.Among them,the inhibition effect was the most obvious in the high-dose group,and the inhibition rate reached 38%,45% and 54% at different time(P<0.05);The cell cycle experiment results showed that compared with the control group,the serum containing YHD significantly changed the cell cycle of MCF-7,blocking the cell cycle in the S phase;The results of DAPI staining showed that,compared with the control group,the serum containing YHD significantly destroyed the MCF-7 nucleus and was dose-dependent;Flow cytometry(Annexin,V-FITC/PI double staining)showed that compared with the control group,the medium dose group and the high dose group with YHD increased the apoptosis rate of MCF-7 cells,and the apoptosis rate was 12.8% and 23.1%,respectively(P<0.05);Western blot analysis showed that compared with the control group,the serum containing YHD reduced the protein expression levels of p38,STAT3,Survivin,bcl-xl and p-stat3 in MCF-7 cells(P<0.05),which were dose-dependent.q RT-PCR assay showed that compared with the control group,the serum containing YHD could reduce the m RNA expression levels of p38,STAT3,Survivin and bcl-xl in Mc F-7 cells(P<0.05),and it was dose-dependent.Conclusion: The YHD drug-containing serum can inhibit the proliferation of MCF-7 cells,and inhibit the expression of p38 by changing cell cycle,destroying the nuclear structure,thus reducing the expression of STAT3,Survivin,bcl-xl and p-stat3,and promoting the apoptosis of cells.The effect of YHD drug-containing serum on MCF-7 cell proliferation may be related to the p38/ STAT3 signaling pathway. |
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