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.
State Key Laboratory of Experimental Hematology, Institute of Hematology &Blood Disease Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China.E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2019 Feb;27(1):233-238. doi: 10.7534/j.issn.1009-2137.2019.01.038.
To explore the effect of damage of bone marrow stroma cells induced by chemotherapeutic drug on the function of normal hematopoitic cells.
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-H2AX was determined by flow cytometry after histone γ-H2AX staining; the colony forming ability of hematopoietic cells was tested by colony formation assay.
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).
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.
探讨化疗药物诱导的骨髓基质细胞损伤对正常造血细胞功能的影响。
采用SA-β-半乳糖苷酶染色后通过流式细胞术检测衰老细胞；运用实时定量聚合酶链反应检测骨髓基质细胞系OP9细胞中一系列分子的表达；通过组蛋白γ-H2AX染色后利用流式细胞术测定γ-H2AX的表达；采用集落形成试验检测造血细胞的集落形成能力。
柔红霉素（DNR）处理后OP9细胞中衰老细胞的百分比是未处理OP9细胞的2.24倍（P<0.05）。与正常OP9细胞相比，DNR处理的OP9细胞中白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的表达水平分别升高了2.73倍（P<0.01）和0.56倍（P<0.01），而N-钙黏蛋白、α-平滑肌肌动蛋白（α-SMA）、血管生成素1（Angpt1）和骨桥蛋白（OPN）的表达水平分别下降了69.54%（P<0.01）、63.90%（P<0.01）、87.41%（P<0.01）和42.78%（P<0.01）。与未处理OP9细胞共培养的正常造血细胞相比，与DNR处理的OP9细胞共培养后，正常造血细胞的集落形成减少了47.10%（P<0.05），同时，正常造血细胞中γ-H2AX+细胞的百分比增加了2.19倍（P<0.05）。
DNR处理后，OP9细胞的衰老细胞数量显著增加；与正常OP9细胞相比，TNF-α和IL-6的表达上调，而α-SMA、Angpt-1和OPN的表达下调。此外，DNR处理的OP9细胞与正常造血细胞共培养后，与正常造血细胞相比，造血细胞的集落形成能力下降，造血细胞的基因组不稳定性增加。
