Bone marrow-derived mesenchymal stromal cells affected acute myeloid leukemia cells via the TSP-1/CD36 pathway
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摘要:
目的 探讨骨髓基质细胞（bone marrow-derived mesenchymal stromal cells，BM-MSCs）对急性髓系白血病（acute myeloid leukemia，AML）细胞影响的作用机制。 方法 构建MLL-AF9过表达诱导的AML小鼠模型，通过PCR比较AML小鼠和野生型小鼠（WT）BM-MSCs内TSP-1的表达差异。通过慢病毒载体使AML小鼠来源的BM-MSCs高表达TSP-1后，与AML细胞行transwell共培养，检测AML细胞表面TSP-1受体CD36及CD47的表达及AML细胞的生长情况。在共培养体系中加入CD36抑制剂N-油酰基硫代琥珀酰亚胺，检测AML细胞增殖、凋亡的变化。 结果 AML小鼠BM-MSCs中TSP-1的表达低于对照组。过表达TSP-1的BM-MSCs与AML 细胞行transwell共培养后AML细胞的生长受到抑制，且AML细胞表面的CD36受体表达升高，但CD47表达无明显差异。在共培养体系中加入CD36抑制剂N-油酰基硫代琥珀酰亚胺后，AML细胞增殖加快，凋亡减少。 结论 TSP-1/CD36信号通路有望成为治疗AML的潜在靶点。 Abstract:Objective To investigate the mechanism of impact of bone marrow-derived mesenchymal stromal cells (BM-MSCs) on acute myeloid leukemia (AML) cells. Methods An overexpressed MLL-AF9-induced AML mouse model was used, and the expression of thrombospondin-1 (TSP-1) was compared between BM-MSCs from AML and wild-type (WT) mice by PCR analysis. TSP-1-overexpressing BM-MSCs were co-cultured with primary AML cells following lentivirus transfection, and the growth of AML cells and expression of TSP-1 receptors (CD36 and CD47) on their surface was detected. The CD36 inhibitor N-oleoylthiosuccinimide was introduced to the co-culture system and AML cell proliferation and apoptosis were observed. Results TSP-1 expression in BM-MSCs derived from AML mice was lower than that in WT mice. Flow cytometry results revealed that CD36 expression increased but CD47 expression remained unchanged in primary AML cells co-cultured with TSP-1-overexpressing BM-MSCs. Furthermore, AML cell growth was inhibited following co-culturing with TSP-1-overexpressing BM-MSCs. The addition of N-oleoylthiosuccinimide increased AML cell proliferation while inhibiting apoptosis. Conclusions TSP-1/CD36 signaling could be a potential therapeutic target for AML. -
图 1 AML 小鼠模型的构建
A: 将过表达MLL-AF9的逆转录病毒载体MSCV-MLL-AF9-PGK-GFP感染B6小鼠来源的Lin-Sca-1+细胞，将感染后GFP+细胞经尾静脉注射至C57小鼠；B、C：建模后14 d解剖小鼠分析脾脏的大小和重量；D、E：建模后14 d解剖小鼠分析肝脏的大小和重量；F：建模后14 d解剖小鼠分析骨髓和脾中白血病细胞的浸润；WT: 野生型小鼠；*：P<0.1；**：P<0.01
图 4 流式检测CD36及CD47的表达水平
A：AML原代细胞表面CD36及CD47的表达情况；B：与过表达TSP-1的BM-MSCs行transwell共培养后的AML细胞表面CD36表达水平升高；C：共培养后CD47的表达未见明显改变）；**：P<0.01
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