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目的探讨膀胱癌MB49细胞来源的外泌体(exosomes)对MDSCs细胞扩增、活化的影响及机制。方法用超滤联合蔗糖重水密度梯度离心法分离纯化膀胱癌MB49细胞分泌的exosomes;电镜形态学分析及Western blotting鉴定;流式细胞仪检测MDSCs比例;ELISA检测VEGF和IL-6;RT-PCR和Western blotting检测cyclin D1、Arg-1 m RNA和蛋白的表达;Western blotting检测p-STAT3和p-NF-κB的变化。结果成功分离纯化得到膀胱癌MB49细胞分泌的exosomes。exosomes可促进MDSCs扩增及活化。实验组MDSCs比例明显升高;Cyclin D1及Arg-1 m RNA和蛋白的表达均上调;p-STAT3和p-NF-κB的表达也上调。结论膀胱癌MB49细胞来源的exosomes能促使MDSCs细胞扩增、活化。研究和分析MDSCs的免疫抑制机制,为疾病的治疗寻找行之有效的新靶点。
Objective To investigate the effect and mechanism of exosomes derived from bladder cancer cell line MB49 on the proliferation and activation of MDSCs. Methods The exosomes secreted by bladder cancer cell line MB49 were isolated and purified by ultrafiltration and sucrose-dextrose gradient centrifugation. The morphology of the exosomes was analyzed by electron microscopy and Western blotting. The proportion of MDSCs was detected by flow cytometry. The levels of VEGF and IL-6 were detected by ELISA. The expression of cyclin D1 and Arg-1 mRNA and protein were detected by Western blotting. The changes of p-STAT3 and p-NF-κB were detected by Western blotting. Results The exosomes secreted by bladder cancer MB49 cells were successfully isolated and purified. Exosomes can promote MDSCs amplification and activation. The proportion of MDSCs in the experimental group was significantly increased; the expressions of Cyclin D1 and Arg-1 mRNA and protein were up-regulated; the expressions of p-STAT3 and p-NF-κB were also up-regulated. Conclusion Exosomes derived from bladder cancer cell line MB49 can promote the proliferation and activation of MDSCs. To study and analyze the immunosuppressive mechanisms of MDSCs and to find new and effective targets for the treatment of diseases.