5-氟尿嘧啶损伤骨髓基质细胞致造血细胞应激诱导性早衰

发布时间：2018-03-22 01:04

本文选题：-氟尿嘧啶　切入点：骨髓基质细胞　出处：《中国实验血液学杂志》2017年04期 　论文类型：期刊论文

【摘要】：目的:探讨抑瘤浓度下的5-氟尿嘧啶(5-fluorouracil,5-FU)对人骨髓基质细胞是否有损伤作用以及该作用对造血细胞的影响。方法:应用CCK-8法测定乳腺癌细胞株MCF-7、结肠癌细胞株HCT-116及人骨髓基质细胞株HS-5对不同浓度5-FU的敏感性。5-FU作用HS-5后,结晶紫染色计数成纤维细胞集落;流式细胞术分析细胞周期;Annexin V/PI双染及Hoechest染色检测细胞凋亡;DCFH-DA法检测胞内活性氧(reactive oxygen species,ROS)水平;ELISA及免疫荧光法检测细胞因子KL、GM-CSF、RANTS、SDF水平。人脐血单个核细胞(human umbilical cord blood mononuclear cell,h UCB-M NC)与HS-5共培养后,台盼蓝染色计数h UCB-M NC;流式细胞术检测细胞周期、ROS水平、CD34+细胞百分率;酶学法检测谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)和超氧化物歧化酶(Superoxide dismutase,SOD)含量;β-半乳糖苷酶染色检测衰老的h UCB-MNC。结果:5-FU 12.5-100μg/ml对M CF-7、HCT-116和HS-5均有增殖抑制作用,且该作用具有浓度依赖性和时间依赖性,其中HS-5对5-FU更为敏感。5-FU作用后HS-5细胞周期阻滞,凋亡率上升,胞内ROS含量显著升高,造血生长因子分泌降低,炎性趋化因子升高。与经5-FU作用的HS-5共培养后,h UCB-MNC数量及CD34+细胞比例降低,G1期阻滞,细胞抗氧化能力降低,胞内ROS含量显著上升,衰老的造血细胞增多。结论:5-FU可导致骨髓基质细胞氧化损伤、分泌生物活性物质改变,诱发造血细胞氧化应激性早衰。
[Abstract]:Objective: to investigate whether 5-fluorouraciline 5-FU (5-fluorouraciline 5-FU) can damage human bone marrow stromal cells (BMSCs) and the effect of 5-fluorouraciline 5-FU (5-FU) on hematopoietic cells. Methods: breast cancer cell line MCF-7, colon cancer cell line HCT-116, and colon cancer cell line HCT-116 were determined by CCK-8 assay. The sensitivity of human bone marrow stromal cell line HS-5 to different concentrations of 5-FU. 5-FU treated HS-5. The colony of fibroblasts was counted by crystal violet staining. Flow cytometry analysis of cell cycle Annexin V/PI double staining and Hoechest staining for detection of apoptosis; Detection of intracellular reactive oxygen species (Ros) by DCFH-DA; Elisa; immunofluorescence assay; detection of the level of cytokine KLGM-CSFRANTSU SDF. Human umbilical cord blood mononuclear cells in human umbilical cord blood mononuclear cells. UCB-M NCC) co-cultured with HS-5, Trypan blue staining counted h UCB-M NC, and flow cytometry was used to detect the percentage of CD34 cells at Ros level. The contents of glutathione peroxidase (Glutathione peroxidase) and superoxide dismutase (SOD) and 尾 -galactosidase (尾 -galactosidase) were detected by enzymatic method. Results the proliferation of MCF-7HCT-116 and HS-5 was inhibited by 5-FU 12.5-100 渭 g/ml. The effect was concentration-dependent and time-dependent. HS-5 was more sensitive to 5-FU. 5-FU could block the cell cycle, increase the apoptosis rate, increase the content of intracellular ROS and decrease the secretion of hematopoietic growth factor. The number of UCB-MNC and the proportion of CD34 cells decreased in G 1 phase after co-culture with 5-FU, and the antioxidant ability of cells decreased, and the content of intracellular ROS increased significantly after co-culture with 5-FU. Conclusion 5-FU can induce oxidative damage of bone marrow stromal cells, secretion of bioactive substances, and induce oxidative stress premature senescence of hematopoietic cells.
【作者单位】：重庆医科大学干细胞与组织工程学研究室组织学胚胎学教研室;
【基金】：国家自然科学基金资助项目(81173398) 重庆市科委基础与前沿研究资助项目(cstc2014jcyj A10001)
【分类号】：R730.5

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