文章摘要
周 丽,陆 化,卢瑞南,王 铃,李建勇,孙幸福.多发性骨髓瘤细胞对血管内皮细胞tPA及PAI⁃1表达的影响[J].南京医科大学学报,2018,(5):600~604
多发性骨髓瘤细胞对血管内皮细胞tPA及PAI⁃1表达的影响
Effect of multiple myeloma cells on the expressions of tPA and PAI⁃1 by the cultured HUVEC
投稿时间:2017-12-12  
DOI:10.7655/NYDXBNS20180506
中文关键词: 多发性骨髓瘤  人脐静脉内皮细胞  组织型纤维蛋白溶酶原激活剂  纤维蛋白溶酶原激活物抑制剂⁃1
英文关键词: multiple myeloma  human umbilical vein endothelial cells  tissue⁃type plasminogen activator  plasminogen activator inhibitor⁃1
基金项目:江苏省中医药局科技项目(L213219);江苏大学医学临床科技发展基金(JLY20160186)
作者单位
周 丽 常州市武进人民医院血液科江苏 常州 213002 
陆 化 南京医科大学第一附属医院血液科江苏 南京 210029 
卢瑞南 南京医科大学第一附属医院血液科江苏 南京 210029 
王 铃 南通市第一人民医院血液科江苏 南通 226001 
李建勇 南京医科大学第一附属医院血液科江苏 南京 210029 
孙幸福 南京市高淳人民医院血液科江苏 南京 211300 
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中文摘要:
      目的:研究多发性骨髓瘤细胞(multiple myeloma cells,MMCs)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)组织型纤维蛋白溶酶原激活剂(tissue?type plasminogen activator,tPA)、纤维蛋白溶酶原激活物抑制剂?1(plasminogen activator inhibitor?1,PAI?1)蛋白表达水平及mRNA表达水平的影响,了解多发性骨髓瘤(multiple myeloma,MM)患者纤溶活性,从而进一步了解MM患者血栓形成的原因及机制。方法:MM细胞株U266与HUVEC共培养24、48、72 h,以同期单独培养的HUVEC、MMCs作为对照,ELISA法检测共培养组和对照组细胞培养上清中tPA、PAI?1的蛋白表达水平,PCR检测HUVEC、MMCs tPA、PAI?1 mRNA的表达水平。结果:无论是ELISA法还是PCR,均未检测到MMCs培养上清中tPA、PAI?1蛋白和mRNA的表达。与同期单独培养的HUVEC相比,ELISA法检测共培养体系中的HUVEC tPA、PAI?1表达水平均升高(P < 0.05);对照组中,PAI?1与tPA的表达水平呈正相关(rs=0.80,P=0.01);HUVEC与MMCs共培养体系中,PAI?1与tPA的表达水平呈正相关(rs =0.88,P=0.002);HUVEC单独培养组中,t?PA、PAI?1表达水平与时间呈正相关(rs=0.90,P=0.001;rs=0.90,P=0.001);共培养组中,t?PA、PAI?1表达水平与时间呈正相关(rs=0.95,P<0.001;rs=0.84,P=0.004);两组中tPA、PAI?1与GAPDH条带灰度值比值无明显差异(P > 0.05)。结论:MMCs可促进HUVEC对tPA及PAI?1的分泌,在保证细胞活力的前提下,随共培养时间延长,两者分泌增加,且PAI?1与tPA的表达呈一定相关性,然而,MMCs对HUVEC tPA及PAI?1 mRNA的表达无明显影响。
英文摘要:
      Objective:To study the influence of multiple myeloma cells(MMCs)on the secretion of tissue?type plasminogen activator(tPA)and tissue?type plasminogen activator(PAI?1)by the cultured human umbilical vein endothelial cells(HUVEC)to confirm the fibrinolytic activity as well as the causes and mechanisms of thrombosis in multiple myeloma(MM). Methods:Human MM cell line U266 was co?cultured with HUVEC for 24 h,48 h and 72 h,with the same period of HUVEC cultured alone as control as well as MMCs. The expressions of HUVEC,MMCs tPA,and PAI?1 mRNA in those groups were detected by polymerase chain reaction(PCR),tPA and PAI?1 protein levels in the supernatant were detected by ELISA. Results:No expression of tPA and PAI?1 protein and mRNA in the culture supernatants of MMCs was detected by both ELISA and PCR. The expression level of tPA protein in the co?culture system by ELISA was significantly(P < 0.05)higher than that in the control group,as well as the PAI?1 protein level(P < 0.05). There was a positive correlation between the PAI?1 and tPA protein level in the control group(rs=0.80,P=0.01)and in the co?culture system(rs=0.88,P=0.002)by ELISA. The levels of t?PA and PAI?1 protein were positively correlated with the time in the HUVEC cultured alone group(rs=0.90,P=0.001;rs=0.90,P=0.001)and the co?culture system(rs=0.95,P<0.001;rs=0.84,P=0.004)significantly by ELISA. The difference of the expression level of tPA mRNA between the two groups was not significant(P > 0.05),as well as the expression level of PAI?1 mRNA(P > 0.05). Conclusion:MMCs can promote the secretion of tPA and PAI?1 protein from HUVEC. On the premise of ensuring the cells viability,the secretion of both proteins increased along with the extension of incubation time,and there was a certain correlation between the tPA and PAI?1 protein level. However,MMCs had no obvious effect on the expression level of tPA and PAI?1 mRNA.
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