Study on synergistic anti-tumor effect and mechanism of anemoside B4 combined with 5-fluorouracil in breast cancer
Objective To examine the combined impact of Anemoside B4(AB4)and 5-fluorouracil(5-Fu)on murine 4T1 breast cancer cells,both in vitro and in vivo,while also investigating the underlying molecular mechanisms.Methods A series of experiments,including cell proliferation,wound healing,matrigel invasion assays,and flow cytometry,were conducted to assess how the combination of AB4 and 5-Fu influences 4T1 cell function and apoptosis in vitro.In vivo,a tumor model was created by subcutaneously injecting 4T1 cells into BALB/c mice to observe the combined effects of AB4 and 5-Fu on tumor growth.HE staining was used to examine changes in tumor tissue microstructure,while flow cytometry analyzed the impact of AB4 and 5-Fu on the mobilization of bone marrow-derived cells(BMDCs).Additionally,qRT-PCR and Western blotting were employed to measure mRNA levels of apoptosis-related factors and the protein expression of angiogenesis-related factors in tumors.Results The combination of AB4 and 5-Fu significantly reduced the viability and the migration and invasion capacity of 4T1 breast cancer cells(P＜0.01).Moreover,the proportion of cells undergoing apoptosis was significantly increased when treated with AB4 and 5-Fu together(P＜0.01);Besides,the combined treatment of AB4 and 5-Fu led to a more significant tumor growth inhibition in 4T1 xenograft mouse model(P＜0.01).The intratumoral microvascular density in 4T1 xenograft was dramatically reduced according to the HE staining results(P＜0.01)and the cell count of BMDCs in peripheral blood of tumor-bearing mice was also reduced(P＜0.01).qRT-PCR analysis shown that AB4 combined with 5-Fu significantly increased the mRNA expression of pro-apoptotic factors and decreased the mRNA levels of anti-apoptotic factors in tumor tissues(P＜0.01),and the protein expression of tumor angiogenesis-related factors was inhibited.Conclusions The combined use of AB4 and 5-Fu effectively inhibited the growth of 4T1 tumor cells in a synergistic manner.This effect is likely due to both the direct inhibition of tumor cell function and the suppression of BMDC mobilization,which in turn reduces tumor angiogenesis indirectly.
万方数据
