急性髓系白血病细胞条件培养液对间充质干细胞的生物学影响  

Effects of conditioned medium of acute myeloid leukemia on biology of mesenchymal stem cells

作　　者：张弛可 王飞清 吴丹 杨波 成金阳 陈娟 唐东昕 刘洋 李艳菊[1,4] Zhang Chike;Wang Feiqing;Wu Dan;Yang Bo;Cheng Jinyang;Chen Juan;Tang Dongxin;Liu Yang;Li Yanju(Guizhou Medical University,Guiyang 550004,Guizhou Province,China;First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,Guizhou Province,China;National&Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique,Guizhou Medical University,Guiyang 550004,Guizhou Province,China;Department of Hematology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China)

机构地区：[1]贵州医科大学,贵州省贵阳市550004 [2]贵州中医药大学第一附属医院,贵州省贵阳市550001 [3]贵州医科大学细胞工程生物医药技术国家地方联合工程实验室,贵州省贵阳市550004 [4]贵州医科大学附属医院血液科,贵州省贵阳市550004

出　　处：《中国组织工程研究》2024年第31期4995-5002,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(31660326),项目负责人:刘洋;国家自然科学基金(82160519),项目负责人:李艳菊;学科优秀后备人才项目(gyfyxkrc-2023-14),项目负责人:李艳菊;贵州省科学技术厅项目(黔科合基础[2016]1019),项目负责人:刘洋;贵阳市科技局项目(2022-4-3-10),项目负责人:王飞清;贵州省高等院校工程研究中心项目(黔教技[2023]037),项目负责人:唐东昕。

摘　　要：背景:目前,关于急性髓系白血病骨髓微环境中的骨髓间充质干细胞生物学功能及分子改变尚不完全清楚。目的:综合生物信息学和细胞实验探究急性髓系白血病患者来源骨髓间充质干细胞的生物学功能改变,并探讨急性髓系白血病细胞条件培养液在其中的作用。方法:从GEO数据集中筛选差异基因进行富集分析,并构建蛋白质互作网络和获取Hub基因。培养急性髓系白血病患者和健康供者的骨髓间充质干细胞,使用急性髓系白血病细胞条件培养液处理健康供者的骨髓间充质干细胞,对各组细胞进行成脂诱导分化、成骨诱导分化、β-半乳糖苷酶染色、细胞周期检测和Hub基因的验证。结果与结论:①从GSE84881中获取急性髓系白血病患者和健康供者的骨髓间充质干细胞基因表达数据,筛选出184个上调基因和140个下调基因。②富集的生物学功能主要包括细胞周期、脂肪细胞分化、细胞代谢过程和MYC通路。根据Degree算法筛选出上调的10个Hub基因和下调的10个Hub基因。③通过体外细胞实验发现,与健康供者的骨髓间充质干细胞相比,急性髓系白血病患者骨髓间充质干细胞表面抗原未发生改变,但其成脂分化能力增强、成骨分化能力减弱、β-半乳糖苷酶阳性细胞数增多、细胞形态改变、细胞周期停滞、LGALS3表达增加以及MYC表达降低。经急性髓系白血病细胞条件培养液培养后,健康供者骨髓间充质干细胞出现相似改变。④结果表明,急性髓系白血病患者骨髓微环境中的间充质干细胞生物学功能发生改变,这为骨髓间充质干细胞与肿瘤的交互作用提供新的见解。BACKGROUND:At present,the biological functions and molecular changes of bone marrow mesenchymal stem cells in the tumor microenvironment of acute myeloid leukemia are still unclear.OBJECTIVE:To explore the changes in the biological function of bone marrow mesenchymal stem cells in acute myeloid leukemia and the role of acute myeloid leukemia conditioned medium by bioinformatics and experiment.METHODS:Differential genes were screened from GEO data sets,and enrichment analysis was performed.The protein-protein interaction network was constructed and the Hub gene was obtained.Bone marrow mesenchymal stem cells from patients with acute myeloid leukemia and healthy donors were cultured.Bone marrow mesenchymal stem cells from healthy donors were treated with acute myeloid leukemia conditioned culture solution.Each group was subjected to the adipogenic differentiation,osteogenic differentiation,staining of β-galactosidase,detection of the cell cycle,and validation of Hub genes.RESULTS AND CONCLUSION:(1)Gene expression data of bone marrow mesenchymal stem cells from acute myeloid leukemia patients and healthy donors were obtained from GSE84881,and 184 up-regulated genes and 140 down-regulated genes were screened.(2)The biological functions of enrichment mainly include cell cycle,adipocyte differentiation,cell metabolism,and MYC pathway.According to the Degree algorithm,10 up-regulated Hub genes and 10 downregulated Hub genes were selected.(3)The cell in vitro experiment found that:compared with the control group,the surface antigen of acute myeloid leukemia mesenchymal stem cells did not change,but it showed enhanced lipid differentiation ability,weakened osteogenic differentiation ability,increasedβ-galactosidase positive cell number,altered cell morphology,arrested cell cycle,increased LGALS3 expression,and decreased MYC expression.Mesenchymal stem cells from healthy donors showed similar changes after being cultured in acute myeloid leukemia conditioned medium.(4)The results show that biological function of mesenchy

关 键 词：急性髓系白血病 骨髓间充质干细胞 条件培养液 GEO数据库 生物信息学 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R394.2