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目的:探讨单核细胞趋化蛋白-1(monocyte chemoatractant protein-1,MCP-1)/趋化因子(C-C模体)受体2[chemokine(C-C motif)receptor 2,CCR2)轴在人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,HUMSCs)向肺癌归巢中的作用。方法:采用组织块培养法从健康新生儿脐带组织中分离HUMSCs并鉴定,构建BALB/c裸鼠皮下移植瘤模型。体外应用Transwell趋化实验、体内应用IVIS Xenogen动物活体成像系统检测HUMSCs是否向肺癌归巢,ELISA法检测肺癌细胞A549培养上清中MCP-1分泌水平,转染shRNA敲低肺癌细胞内MCP-1的表达和用抑制剂RS504393抑制HUMSCs细胞表面的MCP-1受体CCR2后,体内外检测HUMSCs向肺癌归巢能力的改变。结果:成功分离得到HUMSCs并完成鉴定,成功建立BALB/c裸鼠皮下肺癌移植瘤模型。HUMSCs在体内外均能向肺癌归巢(P<0.01),肺癌细胞高表达MCP-1。成功构建稳定低表达MCP-1的肺癌A549细胞系,与对照组相比,shRNA1和shRNA2敲低组趋化HUMSCs的数目明显减少[(80.0±33.0)、(94.0±16.0)vs(167.0±41.0)个,均P<0.05],抑制HUMSCs细胞表面MCP-1受体CCR2后,体内外均显著抑制HUMSCs向肺癌的趋化能力(P<0.05)。结论:MCP-1/CCR2轴促进HUMSCs向肺癌归巢。
OBJECTIVE: To investigate the effect of monocyte chemoatractant protein-1 (MCP-1) / chemokine (CC motif) receptor 2 and CCR2 axis on human umbilical cord The role of human umbilical cord mesenchymal stem cells (HUMSCs) in the homing of lung cancer. METHODS: HUMSCs were isolated from umbilical cord tissue of healthy neonates by tissue culture method and identified. The BALB / c nude mice model was established. In vitro, Transwell chemotaxis assay was used to detect the homing of HUMSCs to lung cancer using IVIS Xenogen imaging system in vivo. The secretion of MCP-1 in the culture supernatant of A549 cells was detected by ELISA and the expression of MCP-1 After inhibiting the MCP-1 receptor CCR2 on the surface of HUMSCs by inhibitor RS504393, the homing ability of HUMSCs to lung cancer cells was detected in vitro and in vivo. Results: The HUMSCs were successfully isolated and identified. The BALB / c nude mice model of subcutaneous lung cancer xenografts was successfully established. HUMSCs could both homing to lung cancer both in vitro and in vivo (P <0.01). MCP-1 was highly expressed in lung cancer cells. Compared with the control group, the number of chemotactic HUMSCs in shRNA1 and shRNA2 knockdown groups was significantly reduced [(80.0 ± 33.0) vs (94.0 ± 16.0 vs 167.0 ± 41.0 ), Both P <0.05]. Inhibition of chemokine MCP-1 receptor CCR2 on HUMSCs significantly inhibited chemotaxis of HUMSCs to lung cancer both in vitro and in vivo (P <0.05). Conclusion: The MCP-1 / CCR2 axis promotes the homing of HUMSCs to lung cancer.