蟾毒灵抑制乏氧耐药细胞诱导的M2型巨噬细胞极化逆转结肠癌耐药  

Bufalin inhibits the polarization of M2 macrophages induced by hypoxic drug-resistant cells and reverses drug resistance in colon cancer

作　　者：唐东豪 王杰[1,2,3] 贾琳琳[1] 江浩鹏 夏琪 陈佳 曹赛雅 陈进宝 司仙科 TANG Donghao;WANG Jie;JIA Linlin;JIANG Haopeng;XIA Qi;CHEN Jia;CAO Saiya;CHEN Jinbao;SI Xianke(Putuo Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China;Department of General Surgery,Shanghai Putuo Central School of Clinical Medicine,Anhui Medical University,Shanghai 200062,China;The Fifth School of Clinical Medicine,Anhui Medical University,Anhui Hefei 230032,China.)

机构地区：[1]上海中医药大学附属普陀医院,上海200062 [2]安徽医科大学上海普陀中心临床学院普外科,上海200062 [3]安徽医科大学第五临床医学院,安徽合肥230032

出　　处：《现代肿瘤医学》2024年第6期987-993,共7页Journal of Modern Oncology

基　　金：国家自然科学基金面上项目(编号:81973625);上海市启明星项目(编号:22YF1441400);安徽医科大学研究生科研与实践创新项目资助(编号:YJS20230098)。

摘　　要：目的:在乏氧微环境中探讨蟾毒灵(bufalin,BU)抑制耐药结肠癌细胞诱导M2型巨噬细胞极化对普通结肠癌细胞的作用。方法:培养基中加入氯化钴(cobalt chloride,CoCl 2)模拟乏氧环境,佛波酯(phorbol 12-myristate 13-acetate,PMA)诱导人源单核细胞THP-1分化为M0巨噬细胞。分别采集乏氧条件下耐药结肠癌细胞、普通结肠癌细胞和BU作用于耐药结肠癌细胞的条件培养基(conditioned medium,CM),然后放入M0巨噬细胞中。通过流式细胞术、Realtime PCR、ELISA实验检测M2型巨噬细胞极化标志因子的表达;运用CCK-8和蛋白免疫印迹(western blot,WB)实验检测耐药结肠癌和普通结肠癌条件培养基诱导M2型巨噬细胞极化后的上清对普通结肠癌的作用。结果:在缺氧环境下,与普通结肠癌细胞相比,耐药结肠癌细胞CM促进M2型巨噬细胞标志物IL-10、TGF-β、CD11b、CD206升高(P<0.01,P<0.05)。极化巨噬细胞的上清液增加了普通结肠癌细胞中P-gp和Bcl-2的表达,同时降低了Bax的表达和对奥沙利铂的敏感性。耐药结肠癌细胞经BU处理后,M2型巨噬细胞标志物IL-10、TGF-β、CD11b、CD206表达降低(P<0.01,P<0.05)。此外,P-gp和Bcl-2的表达减少,而Bax的表达增加,导致对OXA的敏感性增加。结论:乏氧条件下,结肠癌耐药细胞CM促进M2型巨噬细胞极化,蟾蜍灵可通过调节M2型巨噬细胞极化过程逆转结肠癌耐药。Objective:To investigate the effect of bufalin(BU)on common colon cancer cells by inhibiting the polarization of M2 macrophages induced by drug-resistant colon cancer cells in a hypoxic microenvironment.Methods:It involved adding cobalt chloride(CoCl 2)to the medium to simulate a hypoxic environment,and using phorbol 12-myristate 13-acetate(PMA)to induce the differentiation of human monocyte THP-1 into M0 macrophages.The conditioned medium of drug-resistant colon cancer cells under hypoxic conditions,common colon cancer cells,and BU acting on drug-resistant colon cancer cells were collected and then put into M0 macrophages.The expression of M2 macrophage polarization markers was detected using flow cytometry,Realtime PCR,and ELISA experiments.The effects of the supernatant after basal-induced polarization of M2 macrophages on common colon cancer were evaluated using CCK-8 and Western blot experiments.Results:In a hypoxic environment,drug-resistant colon cancer cells CM promoted the increase of M2 macrophage markers IL-10,TGF-β,CD11b,and CD206(P<0.01,P<0.05)compared to common colon cancer cells.The supernatant of polarized macrophages increased the expression of P-gp and Bcl-2 in common colon cancer cells while decreasing the expression of Bax and sensitivity to oxaliplatin.After BU treatment on drug-resistant colon cancer cells,the expression of M2 macrophage markers IL-10,TGF-β,CD11b,and CD206 decreased(P<0.01,P<0.05).Furthermore,the expression of P-gp and Bcl-2 decreased,while the expression of Bax increased,leading to increased sensitivity to OXA.Conclusion:In hypoxic condition,colon cancer drug-resistant cell CM promotes the polarization of M2 macrophages,and bufalin can reverse colon cancer drug resistance by regulating the process of M2 macrophage polarization.

关 键 词：乏氧 蟾毒灵 M2型巨噬细胞 结肠癌 耐药 

分 类 号：R735.3[医药卫生—肿瘤]