miR-15a在多发性骨髓瘤患者外泌体中的表达及外泌体对肿瘤生长的影响  

Expression of miR-15a in Exosomes of Patients with Multiple Myeloma and the Effect of Exosomes on Tumor Cell Proliferation and Apoptosis

作　　者：胡恩赑[1] 金强[2] 方福瑾 陈中新[3] HU En-bi;JIN Qiang;FANG Fu-jin;CHEN Zhong-xin(Bengbu Third Peopled Hospital,Bengbu 233000,China;The First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China)

机构地区：[1]蚌埠市第三人民医院检验科,安徽蚌埠233000 [2]蚌埠市第三人民医院,安徽蚌埠233000 [3]蚌埠医学院第一附属医院,安徽蚌埠233000

出　　处：《肿瘤学杂志》2020年第6期491-495,共5页Journal of Chinese Oncology

摘　　要：[目的]探讨miR-15a在多发性骨髓瘤(MM)患者外泌体中的表达及外泌体对肿瘤发生发展的意义。[方法]选择在我院治疗的MM患者71例及健康志愿者20例,分析miR-15a在血浆外泌体中的表达水平;并以人源MM细胞株MM.1R细胞为实验模型,采用增殖实验观察外泌体对肿瘤细胞增殖的影响,采用流式细胞术检测肿瘤细胞凋亡,采用Western blot检测凋亡通路相关蛋白的表达。[结果]与健康志愿者相比,MM患者的血浆miR-15a水平显著降低(4.45±0.20 vs 12.86±0.63,P<0.001),且与患者临床分期有关(P=0.023)。当使用MM患者来源的外泌体处理MM.1R细胞后,实验组细胞增殖率较对照组提高,在12h、24h、48h时分别提高了14.1%、34.0%、52.7%(P<0.01);而健康志愿者来源的外泌体则使细胞增殖率降低,在12h、24h、48h时分别降低了12.4%、20.8%、27.5%(P<0.01)。流式细胞术结果显示,MM患者来源外泌体抑制了肿瘤细胞凋亡,24h凋亡率减少了3.3%(P=0.005);而健康志愿者来源外泌体促进了肿瘤细胞凋亡,24h凋亡率增加了7.8%(P<0.001)。Western blot检测发现MM组的Bcl-2表达增加,Bax和caspase-3表达下降,与健康志愿者来源的外泌体作用相反。[结论]促凋亡因子miR-15a在MM患者外泌体中的表达水平低于健康人群,且MM患者来源的外泌体具有促进肿瘤发展的作用。[Objective] To investigate the expression levels of miR-15 a in exosomes of patients with multiple myeloma(MM) and the effects of exosomes on apoptosis and proliferation of tumor cells.[Methods] The miR-15 a levels in plasma exosomes of 20 healthy subjects and 71 MM patients were detected. A human MM cell line MM.1 R was used as the experimental model to explore the effect of exosomes on tumor cell proliferation. The apoptosis of tumor cells was detected by flow cytometry,and the expression of proteins related to apoptosis pathways was detected by Western blot. [Results] Plasma miR-15 a levels in MM patients was significantly decreased,compared with healthy subjects(4.45 ±0.20 vs 12.86 ±0.63,P <0.001);and it was associated with the clinical stages(P=0.023). When MM.1 R cells were treated with exosomes from MM patients,the proliferation rate was higher than that of the PBS control group,increased by 14.1%,34.0% and 52.7%after 12 h,24 h and 48 h of treatment,respectively(P<0.01). When MM.1 R cells were treated with exosomes from healthy subjects,the proliferation rate reduced by 12.4%,20.8% and 27.5% after12 h,24 h and 48 h of treatment,respectively(P <0.01). Flow cytometry results showed that cell apoptosis was reduced by 3.3% after 24 h treatment with exosomes from MM patients(P=0.005);cell apoptosis increased by 7.8% 24 h after treatment with exosomes from healthy subjects(P <0.001). Western blot analysis showed that the expression of Bcl-2 increased,Bax and caspase-3 decreased in the MM group,which were contrary to the results of the control group. [Conclusion]The expression of miR-15 a in exosomes of MM patients is down-regulated,and exosomes from MM patients can inhibit apoptosis and promote proliferation of MM cells.

关 键 词：多发性骨髓瘤 外泌体 miR-15a BCL-2 凋亡 

分 类 号：R733.3[医药卫生—肿瘤]