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机构地区:[1]同济大学附属第十人民医院检验科,上海200072
出 处:《同济大学学报(医学版)》2011年第5期41-45,60,共6页Journal of Tongji University(Medical Science)
摘 要:目的研究浆细胞异常增生的多发性骨髓瘤(multiple myeloma,MM)细胞在受到肽聚葡糖(peptidoglycan,PGN)刺激后对细胞增殖的影响。方法利用荧光定量聚合酶链反应(PCR)分析MM U266细胞株TLR2、IL-1β、IL-8、NF-κB、Stat3和TNF蛋白的mRNA水平,与23名正常人单个核细胞(peripheral bloodmononuclear cells,PBMCs)mRNA比较。20μg/ml PGN与U266细胞共培养,分析TLR2和NF-κB蛋白mRNA的变化情况。Western印迹法检测该药对U266细胞NF-κB蛋白的影响。MTT、流式细胞术检测PGN刺激U266后增殖情况。结果与PBMCs相比,TLR2、IL-8、TNF在U266中表达显著降低,而NF-κB表达增加(P<0.05)。经PGN作用后,TLR2、NF-κB mRNA表达均增加,Western印迹法检测NF-κB蛋白表达明显增多。MTTT及细胞周期检测发现,PGN可使细胞由G_0/G_1期向前推进,促进细胞有丝分裂及增殖。结论PGN可能通过激活NF-κB通路促进U266增殖。Objective To investigate the effect of peptidoglycan(PGN) on cell proliferation in multiple myeloma(MM) cell line.Methods The mRNA levels of TLR2,IL-1β,IL-8,NF-κB,Stat3 and TNF were measured by fluorescence quantitative polymerase chain reaction(PCR) in MM U266 cells and the peripheral blood mononuclear cells(PBMCs) of 23 healthy individuals.After U266 cells were cultured with 20μg/ml PGN,the mRNA of TLR2 and NF-κB was analyzed by PCR;and the NF-κB protein was evaluated by Western blotting.In addition,the cell proliferation and cell cycle were analyzed by MTT and Flow cytometry(FCM),respectively.Results Compared with PBMCs,the expressions of TLR2,IL-8,TNF decreased in U266,while the expression of NF-κB increased(P0.05).After treated with PGN,the mRNA level of TLR2 and NF-κB increased,and the expression of NF-κB protein also increased;G_0/G_1 phase was propelled and cell mitosis and proliferation were increased.Conclusion PGN may promote cell proliferation in U266 by activating the NF-κB pathway.
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