Effect Of Marsdenia Tenacissima Injection On The Apoptosis Of Triple-Negative Breast Cancer Cell And The Expression Of SBEM Gene

Objective:Marsdenia tenacissima is the dried stem of the plant of the Apocynaceae family,and is a natural product with inhibitory effects on tumor growth in various types of cancer.SBEM is a secretory protein that is specifically expressed in the mammary glands and salivary glands,and is overexpressed in breast tumors and metastatic lymph nodes.There is a correlation between SBEM expression and the prognosis of breast cancer.This study aims to investigate the effects of different concentrations of MTI on cell proliferation,apoptosis,migration,and SBEM expression in the TNBC cell line MDA-MB-231.The possible mechanism of MTI’s anti-tumor activity against MDA-MB-231 cells was preliminarily explored,providing experimental basis and theoretical support for the clinical application of MTI as a therapeutic drug for TNBC.Methods:1.The CCK-8 method was used to detect the effect of different concentrations of MTI on the proliferation of MDA-MB-231 cells.2.After treatment with MTI,Annexin-FITC/PI staining and flow cytometry were used to detect apoptosis of MDA-MB-231 cells,and a scratch assay was used to detect changes in cell migration ability.3.After treatment with MTI,WB was used to detect the expression of SBEM,BCL-2,and BCL-XL proteins in MDA-MB-231 cells.4.After treatment with MTI,RT-q PCR was used to detect changes in SBEM m RNA expression levels in the cells.Results:The CCK-8 experiment results showed that different concentrations of MTI effectively inhibited the proliferation of MDA-MB-231 cells,and there was a statistically significant difference in cell viability between groups(P<0.01).The inhibitory effect of MTI on the proliferation of MDA-MB-231 cells was concentration-dependent.Flow cytometry analysis revealed that compared with the negative control group,the apoptosis rate of MDA-MB-231 cells increased after MTI treatment,and the apoptosis rates at MTI concentrations of 0mg/ml,10mg/ml,20mg/ml,40mg/ml,and 60mg/ml were(4.05±0.08)%,(6.30±0.13)%,(12.28±0.85)%,(22.80±1.76)%,and(34.06±1.09)%,respectively.The pro-apoptotic effect of MTI at concentrations of 20mg/ml,40mg/ml,and 60mg/ml had a statistically significant difference(P<0.001).RT-q PCR and WB experiments revealed that the apoptosis-related proteins BCL-2 and BCL-XL were inhibited in MDA-MB-231 cells after treatment with different concentrations of MTI,and the differences were statistically significant compared with the negative control group(P<0.05).In addition,the expression inhibition effect of SBEM was not significant at an MTI concentration of 10mg/ml(P>0.05),while it was significant in all other groups(P<0.05).Different concentrations of MTI significantly inhibited SBEM m RNA(P<0.05),and the differences between the concentration groups were statistically significant(P<0.05).Conclusions:This study found that MTI has significant anti-tumor activity through in vitro cell experiments by reducing the proliferation activity of MDA-MB-231 cells,promoting cell apoptosis,and inhibiting cell migration.Based on the experimental results,MTI may affect the proliferation,apoptosis,and migration of MDA-MB-231 cells by downregulating BCL-2,BCL-XL,and reducing SBEM expression.