白头翁皂苷B_(4)与5-氟尿嘧啶联用抗乳腺癌作用及机制  

Study on synergistic anti-tumor effect and mechanism of anemoside B_(4) combined with 5-fluorouracil in breast cancer

作　　者：张琦 田冲冲 ZHANG Qi;TIAN Chongchong(College of Pharmacy,Jiangsu Medical College,Yancheng 224005,China)

机构地区：[1]江苏医药职业学院药学院,江苏盐城224005

出　　处：《药物评价研究》2024年第11期2574-2584,共11页Drug Evaluation Research

基　　金：江苏医药职业学院自然科学基金研究一般项目(20214107);江苏高校哲学社会科学研究一般项目资助(2022SJYB2075);江苏省盐城市卫生健康委员会2023年度医学科研立项项目(YK2023032,YK2023097)。

摘　　要：目的从体内外水平探究白头翁皂苷B_(4)(AB_(4))与5-氟尿嘧啶(5-Fu)联合用药对小鼠4T1乳腺癌细胞的作用及其分子机制。方法体外分别采用细胞增殖抑制实验、细胞创伤愈合实验、Transwell小室侵袭实验、流式细胞分析以及实时荧光定量PCR法(qRT-PCR)检测AB_(4)与5-Fu联合对4T1细胞功能和凋亡的影响;体内借助4T1乳腺癌荷瘤小鼠模型,观察AB_(4)与5-Fu联合对移植瘤生长的影响;通过苏木精-伊红(HE)染色法分析联合用药对肿瘤组织形态学和微血管密度的影响;流式细胞术检测AB_(4)联合5-Fu对骨髓源性细胞(BMDCs)动员的影响;最后,通过qRT-PCR和Western blotting技术分析对肿瘤组织中相关因子mRNA及蛋白表达的影响。结果体外实验证实AB_(4)联合5-Fu显著降低4T1乳腺癌细胞活力,并且抑制其迁移和侵袭能力,同时可诱导其凋亡(P<0.01);体内实验表明AB_(4)联合5-Fu显著抑制4T1移植瘤的生长(P<0.01),减少肿瘤组织内微血管密度以及外周循环血液中促血管新生BMDCs的细胞计数(P<0.01),还能显著上调肿瘤组织中促凋亡因子的mRNA水平及降低抗凋亡因子的mRNA水平(P<0.01),同时抑制肿瘤血管新生相关因子蛋白的表达(P<0.01)。结论AB_(4)与5-Fu联合使用可以协同抑制4T1肿瘤细胞的生长。机制可能与其对肿瘤细胞功能的直接抑制作用有关,同时也与其对BMDCs动员的抑制相关,这间接减少了肿瘤血管新生。Objective To examine the combined impact of Anemoside B_(4)(AB_(4))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 AB_(4) 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 AB_(4) and 5-Fu on tumor growth.HE staining was used to examine changes in tumor tissue microstructure,while flow cytometry analyzed the impact of AB_(4) 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 AB_(4) 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 AB_(4) and 5-Fu together(P<0.01).Besides,the combined treatment of AB_(4) 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 AB_(4) 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 AB_(4) and 5-Fu effectively inhibited the growth of 4T1 tumor cells in a synergistic manner.This effect is likely due to b

关 键 词：白头翁皂苷B_(4) 5-氟尿嘧啶 乳腺癌 协同作用 血管新生 凋亡 

分 类 号：R285.5[医药卫生—中药学]