Preliminary Study On Efficacy Evaluation And Related Mechanisms Of Dasatinib And Docetaxel On Triple-Negative Breast Cancer Mice Models

Objective:To study the effects and mechanisms of dasatinib and docetaxel on inhibiting the growth of triple negative breast cancer in vivo.To investigate whether dasatinib can replace traditional chemotherapy,or combined with traditional chemotherapy to improve the treatment of advanced triple negative breast cancer.Methods:Human breast cancer MDA-MB-231 cells were selected and cultured in RPMI-1640 medium and cultured at 37℃ in a volume fraction of 5%CO2 incubator.The logarithmic growth phase of MB-231 cells after trypsinization,injected in the back of the right subcutaneous NPG mice(0.2ml),6-8 days to form tumors.The tumor-forming mice were randomly divided into four groups,namely Dasatinib and docetaxel combination group,Dasatinib group,Docetaxel group and Control group were given different drugs for treatment.The body weight and tumor size were measured during the treatment of mice,and the tumor growth curves were drawn Tumor inhibition rate(TIR)of the transplanted tumor in each group was measured.After 3 weeks of treatment,the mice in each group were sacrificed by cervical dislocation,the breast tumor tissues were completely dissected,and the histopathology was confirmed by HE staining.The remaining tumor tissues were prepared into single cell suspensions.Flow cytometry was performed on ALDEFLUOR Method to determine the intra-tumor breast cancer stem cells in each group of mice.The total RNA of the remaining single cell suspension was extracted and sequenced.The quantitative and differential analysis of gene expression in each sample was determined,and the enrichment analysis of GO and KEGG was performed.Results:1、The body weight of mice in all drug intervention groups changed.Overall,there was no significant difference between the two groups(P>0.05).There was significant statistical difference between the two groups in the Control group and Docetaxel group(P<0.01).2、There was no significant difference between the other groups(P>0.5).The tumor volume of four groups of mice at different times were compared.The tumor volume of the four groups of mice treated by 5 days,10 days,15 days and 20 days were compared.The comparison of the two groups showed significant differences(P<0.01)Dasatinib + Docetaxel group<Dasatinib group<Docetaxel group<Control group.The tumor inhibition rates of the three groups were(85.72±1.03)%,(65.36±3.58)%,(51.01±4.72)%,respectively.Dasatinib + Docetaxel group>Dasatinib group>Docetaxel group,the overall comparison,any pairwise comparison was statistically significant(P<0.01).3、A total of 17,464 genes were detected in the transcriptome sequencing,among which SAA1,TEM183B,DNER,and LDLRAD2 genes were significantly up-regulated.Conclusion:1、dasatinib regardless of single-drug or combination chemotherapy for the late-stage triple negative breast cancer mice showed a significant anti-tumor effect.2、Dasatinib for inhibiting breast cancer stem cells in vivo showed a strong role,combined application of docetaxel can increase the proportion of cancer stem cells in breast cancer cells.