人骨髓来源间充质干细胞在肿瘤微环境中上调IL-17A水平激活PI3K/Akt通路促进弥漫大B细胞淋巴瘤生长和化疗耐药  

Human Bone Marrow-derived Mesenchymal Stem Cells Promote Growth and Drug-resistance of Diffuse Large B-cell Lymphoma in Tumor Microenvironment by Up-regulating IL-17A Expression via Activating PI3K/Akt Pathway

作　　者：钟伟杰[1] 李庆山[1] 李康保[1] 许昕[1] 魏亚明[1] 朱园[1] 朱志刚[1] ZHONG Wei-jie;LI Qing-shan;LI Kang-bao;XU Xin;WEI Ya-ming;ZHU Yuan;ZHU Zhi-gang((kiangzhou First People's Hospital,Guangzhou 510180,China)

机构地区：[1]广州市第一人民医院,广东广州510180

出　　处：《中国肿瘤》2020年第5期379-390,共12页China Cancer

基　　金：广东省自然科学基金项目(2018A0303130167);广东省医学科学技术研究基金项目(A2018258);广州市卫生和计划生育科技项目(20181A011011)。

摘　　要：[目的]探讨影响弥漫大B细胞淋巴瘤(DLBCL)生长和化疗耐药的机制,并分析其中的信号通路。[方法]分别采用CCK-8法检测DLBCL细胞株(SU-DHL-2和SU-DHL-4细胞)的增殖,用qRTPCR和ELISA法检测白细胞介素(IL)-17A的表达,用流式细胞术分析Th17和Treg细胞的表达,用PI/Annexin V法检测细胞凋亡,用基因芯片分析、生物信息分析和蛋白质印迹法验证IL-17A介导的DLBCL生长的信号通路。[结果 ]人骨髓来源间充质干细胞(hBMSCs)联合外周血单个核细胞(PBMCs)在体外促进DLBCL细胞株的增殖。hBMSCs升高PBMCs中Th17和Treg细胞比例,上调IL-17A和TGF-β水平。hBMSCs显著上调PBMCs中RORγt、Foxp3、IL-17A和TGF-βmRNA的相对表达,增加共培养上清液中IL-17A和TGF-β蛋白的水平。IL-17A通过抑制自发或药物诱导的凋亡促进DLBCL细胞增殖和化疗耐药。外源性IL-17A促进了SU-DHL-4细胞的增殖,降低了自发或利妥昔单抗诱导的SU-DHL-4细胞凋亡,而aIL-17A则消除了这些作用。IL-17A上调细胞周期蛋白D2激活PI3K/Akt信号通路。[结论] hBMSCs在肿瘤微环境中上调IL-17A水平,激活PI3K/Akt通路,促进DLBCL生长和化疗耐药。[Purpose]To investigate the effect of human bone marrow-derived mesenchymal stem cells(hBM-MSCs) on growth and drug-resistance of diffuse large B-cell lymphoma(DLBCL) and its mechnism.[Methods] DLBCL SU-DHL-2 and SU-DHL-4 cells were treated with hBMSCs and peripheral blood mononuclear cells(PBMCs) in vitro. CCK-8 assay was used to detect the proliferation of SU-DHL-2 and SU-DHL-4 cells;qRT-PCR and ELISA were used to study the expression of IL-17A,flow cytometry was used to analyze the percentages of Th17cells and Treg cells;microarray analysis,bioinformatics analysis and Western blot were used to analyze the pathways of IL-17A mediated DLBCL growth. [Results] The h BMSCs promoted the proliferation of DLBCL SU-DHL-2 and SU-DHL-4 cells in vitro. hBMSCs induced PBMCs differentiation into Th17and Treg cells,and increased IL-17A and TGF-β levels in the co-culture supernatants. The hBMSCs significantly up-regulated the relative expressions of RORγt,Foxp3,IL-17A and TGF-β mRNA in PBMCs,increased the levels of IL-17A and TGF-β proteins in the supernatants. IL-17A promoted the growth and drug-resistance of DLBCL cells by inhibiting spontaneous or drug-induced apoptosis. Exogenous IL-17A promoted SU-DHL-4 cell proliferation and inhibited SU-DHL-4 cell spontaneous or drug-induced apoptosis,whereas IL-17A abolished these effects. IL-17A up-regulated cyclin D2 and activated the PI3K/Akt signaling pathway.[Conclusion] The hBMSCs promote the growth and drug-resistance of DLBCL by elevating IL-17A level via PI3K/Akt pathway in the tumor microenvironment.

关 键 词：间充质干细胞 白细胞介素-17A PI3K/AKT 弥漫性大B细胞淋巴瘤 

分 类 号：R73-3[医药卫生—肿瘤] R733[医药卫生—临床医学]