| 尹芳芳,许磊晶,贺美娟,王 悍.甲氨蝶呤诱导巨噬细胞M1极化促进骨肉瘤细胞凋亡的实验研究[J].,2020,(11):2006-2011 |
| 甲氨蝶呤诱导巨噬细胞M1极化促进骨肉瘤细胞凋亡的实验研究 |
| Experimental Study of Methotrexate Induced M1 Polarization of Macrophages to Promote Apoptosis of Osteosarcoma Cells |
| 投稿时间：2020-02-27 修订日期：2020-03-21 |
| DOI：10.13241/j.cnki.pmb.2020.11.002 |
| 中文关键词: 甲氨蝶呤 巨噬细胞 骨肉瘤 |
| 英文关键词: Methotrexate Macrophage Osteosarcoma |
| 基金项目:国家自然科学基金项目（81871400）；上海市优秀技术带头人计划项目（2017XD1424200）；上海市卫生计生系统优秀学科带头人计划项目（2017BR038） |
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| 中文摘要: |
| 摘要 目的：探讨甲氨蝶呤诱导巨噬细胞分化情况和分化后巨噬细胞对骨肉瘤细胞凋亡的影响。方法：采用甲氨蝶呤刺激小鼠单核巨噬细胞RAW264.7细胞24小时后，使用流式、免疫荧光等技术检测M1型巨噬细胞标志物CD86、诱导型一氧化氮合酶（Inducible nitric oxide synthase，iNOS）的表达量，并以脂多糖(Lipopolysaccharide, LPS)诱导巨噬细胞极化为阳性对照，未处理细胞为阴性对照，评估甲氨蝶呤的诱导效果。将经甲氨蝶呤刺激的巨噬细胞与骨肉瘤细胞K7共培养，使用流式技术检测骨肉瘤细胞K7凋亡程度。结果：一定剂量的甲氨蝶呤作用于小鼠单核巨噬细胞后，可以显著上调M1型巨噬细胞的标志物CD86、iNOS，上调程度与LPS组相当。与脂多糖诱导巨噬细胞极化类似，甲氨喋呤可以激活NF-κB。经甲氨蝶呤刺激后的巨噬细胞可以促进骨肉瘤细胞的凋亡。结论：甲氨蝶呤诱导向M1型分化的巨噬细胞可以促进骨肉瘤细胞的凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the differentiation in methotrexate-induced macrophages and the effect of the macrophages on apoptosis of osteosarcoma cells. Methods: Macrophages were stimulated with methotrexate for 24 hours. The expressions of CD86 and inducible nitric oxide synthase, which are markers of M1-like macrophages, were respectively detected by flow cytometry and immunofluorescence. To evaluate the effect of methotrexate on the polarization of macrophages, lipopolysaccharide, LPS, was adopted as a positive control, and untreated cells were used as a negative control. The macrophages, stimulated with MTX, were co-cultured with osteosarcoma cells to assess the effects of the macrophages on the apoptosis of osteosarcoma cells. The rates of apoptosis in osteosarcoma cells were detected by Flow cytometry. Results: After co-cultured with methotrexate, the expression of M1-like macrophage' markers, such as CD86 and iNOS in macrophages was significantly up-regulated, and the degree of upregulation is comparable to that of the LPS group. Similar to LPS-induced macrophage' polarization, methotrexate can cause NF-κB activation. Macrophages stimulated by methotrexate can increase the apoptosis rate of osteosarcoma cells. Conclusion: Methotrexate can induce macrophages to differentiate into M1 like and differentiated macrophages can promote the apoptosis of osteosarcoma cells. |
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