骨髓间充质干细胞条件培养液对多发性骨髓瘤细胞增殖的影响  

Effect of bone marrow mesenchymal stem cell conditioned medium on the proliferation of multiple myeloma cells

作　　者：黄美 王飞清 梁惠玲 杨姁 赵嘉宁 王琨 刘燕青 周媛 王季石[1,3] 李艳菊 刘洋[2,4] Huang Mei;Wang Feiqing;Liang Huiling;Yang Xu;Zhao Jianing;Wang Kun;Liu Yanqing;Zhou Yuan;Wang Jishi;Li Yanju;Liu Yang(Guizhou Medical University,Guiyang 550004,Guizhou Province,China;First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,Guizhou Province,China;Department of Hematology,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou Province,China;National&Guizhou Joint Engineering Laboratory for Cell Engineering and Biomedicine Technique,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]贵州医科大学附属医院血液科,贵州省贵阳市550004 [4]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

出　　处：《中国组织工程研究》2022年第19期3024-3029,共6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;中国博士后科学基金(2018M640938),项目负责人:刘洋;中国博士后科学基金(43XB3794XB),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;贵州省科学技术厅项目(黔科合LH字[2016]7505),项目负责人:刘燕青;贵州省科学技术厅项目(黔科合LH字[2017]7140号),项目负责人:王飞清;贵州省教育厅拔尖人才项目(黔教合KY[2016]074),项目负责人:刘洋;贵州省中医药管理局项目(QZYY-2018-062),项目负责人:周媛;贵中医科院内[2019]45号,项目负责人:王琨。

摘　　要：背景:近年来,骨髓微环境与多发性骨髓瘤细胞生长、存活及耐药性的关系成为科研人员研究热点,骨髓间充质干细胞条件培养液干预多发性骨髓瘤细胞株的实验鲜有报道。目的:探讨骨髓间充质干细胞条件培养液对多发性骨髓瘤细胞增殖的影响。方法:经Ficoll密度梯度离心法及贴壁纯化健康供者骨髓来源间充质干细胞,收集第3-5代骨髓间充质干细胞的上清培养基,超滤离心管制备浓缩骨髓间充质干细胞条件培养液,制备含10%人骨髓间充质干细胞条件培养液的1640完全培养基,诱导多发性骨髓瘤RPMI8226和U266细胞株培养48h,MTT法、EdU荧光染色检测细胞增殖能力;流式细胞仪和PI荧光染色检测细胞凋亡情况;Real-time PCR和Western blot检测细胞中BCL-2、BTK mRNA、蛋白表达水平。结果与结论:骨髓间充质干细胞条件培养液能促进多发性骨髓瘤细胞株的增殖(P<0.05),抑制多发性骨髓瘤细胞的凋亡(P<0.05),与BCL-2、BTK的mRNA和蛋白高表达密切相关(P<0.05)。结果表明,骨髓间充质干细胞条件培养液可促进多发性骨髓瘤细胞增殖。BACKGROUND:In recent years,relationship between the bone marrow microenvironment and the growth,survival and drug resistance of multiple myeloma cells has become a research hotspot for researchers.Few studies have evaluated the effects of bone marrow mesenchymal stem cells conditioned culture medium on multiple myeloma cell lines.OBJECTIVE:To investigate the effect of bone marrow mesenchymal stem cell conditioned medium on the proliferation of multiple myeloma cell lines.METHODS:Ficoll density gradient centrifugation and adherent method were used to purify bone marrow mesenchymal stem cells from healthy donors.The supernatants of 3-5 generations of bone marrow mesenchymal stem cells were collected in the culture medium.Ultrafiltration centrifuge tube was used to prepare bone marrow mesenchymal stem cell conditioned medium and to prepare 1640 complete medium containing 10% human bone marrow mesenchymal stem cell conditioned medium so as to induce the culture of multiple myeloma cell lines RPMI8226 and U266 for 48 hours.Cell proliferation was detected by MTT assay and EdU fluorescence staining.Cell apoptosis was detected by flow cytometry and PI fluorescence staining.Real-time PCR and western-blot assay were used to detect mRNA and protein levels of multiple myeloma cell lines BCL-2 and BTK.RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cell conditioned medium could promote the proliferation of multiple myeloma cell lines(P<0.05),reduce the apoptosis rate of cancer cells(P<0.05)and was associated with high mRNA and protein expression of BCL-2 and BTK(P<0.05).The results showed that the bone marrow mesenchymal stem cell conditioned medium promoted the proliferation multiple myeloma cell lines.

关 键 词：骨髓间充质干细胞 多发性骨髓瘤 条件培养液 增殖 凋亡 

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