| Breast cancer,as one of the most common malignant tumors in women,accounts for 30% of women's cancers.The prevalence of breast cancer in China has increased year by year,and new morbidity and mortality account for approximately 12.2% and 9.6% of the world's total morbidity.As a traditional treatment method for breast cancer,surgical treatment,chemotherapy and radiotherapy are accompanied by serious side effects such as hair loss,nausea,vomiting,thrombosis and bone marrow suppression.Therefore,it is urgent to develop new anticancer drugs with low toxicity and side effects.Over the years,with the continuous deepening of research on Chinese medicine,more and more people have discovered that traditional Chinese medicine not only has significant therapeutic effects in preventing and treating cancer,but also has little toxic and side effects.Many Asian countries have also formed a complete theory,diagnosis and treatment systems,so its applications are becoming more widespread worldwide.Traditional Chinese medicine theory believes that the occurrence of breast cancer is affected by emotional factors such as qi deficiency,blood stasis and phlegm turbidity,among which liver qi stagnation is the first.Based on the traditional Chinese medicine theory of ‘smoothing liver,nourishing the blood and invigorating spleen to cure the root cause;soften hardness and dissolve lump,heat-clearing and detoxifying to cure the symptoms',Shuganhuazheng Paste is an empirical prescription developed by Professor Zhang Yue for the treatment of breast cancer.It has satisfactory clinical effect,but its application is limited due to its unclear mechanism.Therefore,we will study the effects of Shuganhuazheng Paste on the proliferation and apoptosis of MDA-MB-231 cells and the related mechanisms.Objects:By studying the proliferation and apoptosis of MDA-MB-231 cells,we will demonstrate the inhibitory effect of Shuganhuazheng Paste on breast cancer,and find an inhibitory mechanism of Shuganhuazheng Paste on breast cancer,in order to provide an important experimental basis for the clinical application of Shuganhuazheng Paste in the treatment of breast cancer.Methods:The cell viability and proliferition were determined by CCK8 assay and flat clones formation assay at different concentrations of Shuganhuazheng Paste;Hoechst 33342 staining,flow cytometry and JC-1 staining were used to detect the effect of Shuganhuazheng Paste on MDA-MB-231 cells apoptosis;Western blot was used to detect the effects of Shuganhuazheng Paste on the expression of Bax,Bcl-2 and cleaved-caspase 3;Real-time quantitative PCR was used to detect the effect of Shuganhuazheng Paste on the expression of PTEN mRNA;Western blot was used to detect the effect of Shuganhuazheng Paste on the expression of PTEN protein in MDA-MB-231 cells and the activation of its' downstream signaling molecule AKT.Results:First,compared with the control group,the cell viability of the Shuganhuazheng Paste group was significantly reduced(P<0.001?0.0001),80mg/mL concentration is the most obvious(P<0.0001);Secondly,Shuganhuazheng Paste increased the intracellular content of pro-apoptotic protein Bax(P<0.01?0.001),and reduced the intracellular content of anti-apoptotic protein Bcl-2(P<0.05?0.01),and also could increase the activation of caspase 3(P<0.01?0.001),Eventually promoted the apoptosis of MDA-MB-231 cells.At last,it was found that Shuganhuazheng Paste could inhibit the activation of AKT by promoting the expression of PTEN in MDA-MB-231 cells.Conclusions:(1)Shuganhuazheng Paste can inhibit the proliferation of MDA-MB-231 cells.(2)Shuganhuazheng Paste can promote the apoptosis of MDA-MB-231 cells.(3)Shuganhuazheng Paste may play a role in inhibiting proliferation and romoting apoptosis of MDA-MB-231 cells by up-regulating the expression f PTEN,reducing intracellular levels of p-AKT and affecting the PI3K/AKT ignaling pathway.(4)Shuganhuazheng Paste can be used for the treatment of breast cancer. |
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