化疗引起的骨髓基质细胞损伤对正常造血细胞的影响  

Effect of Chemotherapeutic Drug-Induced Damage of Bone Marrow Stroma Cells on Normal Hematopoietic Cells

作　　者：李艺辉 刘喆 李欢[1] 徐颖茜 邢海燕[1] 田征[1] 唐克晶[1] 王敏[1] 饶青[1] LI Yi-Hui;LIU Zhe;LI Huan;XU Ying-Xi;XING Hai-Yan;TANG Ke-Jing;TIAN Zheng;WANG Min;RAO Qing(State Key Laboratory of Experimental Hematology,Institute of Hematology & Blood Disease Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College,Tianjin 300020,China)

机构地区：[1]中国医学科学院北京协和医学院血液学研究所血液病医院实验血液学国家重点实验室,天津300020

出　　处：《中国实验血液学杂志》2019年第1期233-238,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(NO.81770106.);中国医学科学院医学与健康创新工程项目(2016-I2M-1-017)

摘　　要：目的:检测化疗药物柔红霉素(DNR)对小鼠骨髓基质细胞细胞系OP9的损伤作用,探索受损伤后的OP9细胞对正常造血细胞功能的影响。方法:β-gal底物探针孵育后流式细胞术检测细胞衰老;实时荧光定量PCR检测OP9细胞的分子表达;流式细胞技术检测组蛋白γ-H2AX的表达;集落形成实验观察造血细胞集落形成能力的变化。结果:DNR处理后的OP9衰老细胞比例为正常OP9衰老细胞的2.24倍(P<0.05)。同正常OP9细胞相比,OP9细胞经DNR处理后IL-6和TNF-α的表达分别上升了2.73倍(P<0.01)和0.56倍(P<0.01),而神经型钙粘附蛋白(N-cadherin),α平滑肌机动蛋白(α-SMA),重组人血管生成素1(Angpt1),骨桥蛋白(OPN)的表达分别下降了69.54%(P<0.01),63.90%(P<0.01),87.41%(P<0.01)和42.78%(P<0.01)。同正常OP9细胞相比,DNR处理的OP9细胞同正常造血细胞共培养之后,造血细胞的集落形成能力降低了47.10%(P<0.05)。流式细胞术检测显示,正常造血细胞γ-H2AX阳性率增加了2.19倍(P<0.05)。结论:DNR处理后OP9细胞与正常OP9细胞相比,衰老细胞的数量明显增多;TNF-α和IL-6表达升高,N-cadherin,α-SMA, Angpt1和OPN的表达有所下降。与正常OP9细胞相比,DNR处理OP9细胞与正常造血细胞共培养之后,造血细胞的集落形成能力降低,造血细胞基因组不稳定性增加。Objective: To explore the effect of damage of bone marrow stroma cells induced by chemotherapeutic drug on the function of normal hematopoitic cells. Methods: Senescence cells were detected by flow cytometry after SA-β-gal staining;real-time PCR was used to detect the expression of a serial molecules in bone marrow stromal cell line OP9 cells;the expression of γ-H2 AX was determined by flow cytometry after histone γ-H2 AX staining;the colony forming ability of hematopoietic cells was tested by colony formation assay. Results: The percentage of senescence cells in OP9 cells after DNR treatment was 2.24 times as much as that in untreated OP9 cells(P<0.05). Compared with normal OP9 cells, the expression levels of IL-6 and TNF-alpha in DNR-treated OP9 cells increased by 2.73 times(P<0.01) and 0.56 times(P<0.01), and the expression levels of N-cadherin, alpha smooth muscle actin(alpha-SMA), angiopoietin1(Angpt1) and osteopontin(OPN) decreased by 69.54%(P<0.01),63.90%(P<0.01),87.41%(P<0.01)and 42.78%(P<0.01)respectively. After the co-culture with DNR-treated OP9 cells, the colony formation of normal hematopoietic cells decreased by 47.10% than that co-cultured with untreated OP9 cells(P< 0.05), meanwhile, the percentage of γ-H2AX+ cells in normal hematopoietic cells increased by 2.19 times(P<0.05). Conclusion: After treatment with DNR, the senescence cell number of OP9 cells sgnificantly increases;the expression of TNF-α and IL-6 is up-regulated, while the expression of α-SMA, Angpt-1 and OPN is down-regulated as compared with normal OP9 cells. In addition, after co-culture of DNR-treated OP9 cells with normal hematopoietic cells, the colony formation ability of hematopoietic cells decreases and the genome instability of hematopoietic cells increases as compared with normal hematopoietic cells.

关 键 词：柔红霉素 OP9细胞 造血细胞细胞衰老 

分 类 号：R331.2[医药卫生—人体生理学] R730.53[医药卫生—基础医学]