神经元特异性烯醇化酶和U266细胞株对破骨样细胞作用的研究  

Study on effect of NSE and U266 cell line on osteoclast-like cells

作　　者：袁忠涛[1] 卿晨[2] 李晓进[3] 张学美[1] 史明霞[1] 黄颖[1] 李惠民[1] 

机构地区：[1]昆明医科大学第一附属医院血液科,650032 [2]昆明医科大学药学院暨重点实验室,650500 [3]昆明医科大学第四附属医院血液科,650021

出　　处：《重庆医学》2016年第10期1309-1312,共4页Chongqing medicine

基　　金：云南省应用基础研究面上项目(2007C0037R)

摘　　要：目的探讨神经元特异性烯醇化酶(NSE)和人多发性骨髓瘤细胞U266对人破骨样细胞(OLC)功能的影响。方法采用正常人外周血单个核细胞加用细胞核因子κB受体激治蛋白配体(RANKL)+巨噬细胞集落刺激因子(M-CSF)的方法诱导培养OLC;实验分3组,NSE组:将6孔培养板中培养14d的OLC,分别加入100ng/mL重组人NSE培养24、48、72h;共培养组:将6孔Transwell小室下室中培养14d的OLC,各上室分别加入1×105/孔U266细胞进行共培养24、48、72h;对照组:单独培养的OLC。通过实时荧光定量PCR比较NSE和U266细胞株对OLC的RANKL、扩胃蛋白(OPG)、IL-6和抗酒石酸酸性磷酸酶(TRAP)mRNA转录水平的影响。结果共培养组、NSE组和对照组RANKL、OPG、IL-6mRNA在OLC均不表达;与对照组相比,共培养组、NSE组的TRAP mRNA表达水平升高,差异有统计学意义(P<0.01);TRAP mRNA表达水平的升高尤其以48h和72h明显。结论本试验中OLC表达TRAP,NSE是促进骨髓瘤骨病中OLC分化和成熟的因素,提示NSE能增强OLC的活性。Objective To explore the functions of neuron-specific enolase（NSE）and human multiple myeloma U266 cells on osteoclast-like cells（OLC）function.Methods Normal human peripheral blood mononuclear cells were induced and cultured by adding RANKL and M-CSF to get OLC;the experiment was divided into 3groups,the NSE group：OLC were cultured in the 6-well culture plate for 14 dand added with 100ng/mL recombinant human NSE to culture for 24,48,72h;the co-culture group：OLC were cultured in the lower well of 6-well Transwell chamber for 14 d,then added with 1×105/well U266 cells in each upper well and conducted the co-culture for 24,48,72h;the control group：OLC were cultured alone.The influences of NSE and U266 cell line on RANKL,OPG,IL-6and TRAP mRNA transcriptional level of OLS were compared by using real-time fluorescent quantitative PCR.Results RANKL,OPG,IL-6mRNA had no expression on OLC in the co-culture group,NSE group and control group;compared with control group,the TRAP mRNA expression level in the co-culture group and the NSE group was increased,the difference was statistically significant（P〈0.01）;the increase of TRAP mRNA expression level was obvious especially at 48,72 h.Conclusion OLC expressing TRAP and NSE may be one of the factors for promoting OLC differentiation and maturation in myeloma bone disease,prompting that NSE could increase the OLC viability.

关 键 词：磷酸丙酮酸水合酶 骨髓瘤骨病 破骨细胞 酸性磷酸酶 

分 类 号：R733.3[医药卫生—肿瘤]