骨髓间充质干细胞与骨髓瘤细胞共培养时EphB4/ephrinB2表达异常  

The EphB4/ephrinB2 axis is dysregulated in bone marrow derived mesenchymal stem cells(MSCs) after communication with myeloma cell lines

作　　者：王欢[1] 李皎[1] 朱彦[1] 陆化[2] 童珊珊[3] 余先球[1] 王丽霞[1] 汤郁[1] 费小明[1] 

机构地区：[1]江苏大学附属医院血液科,江苏镇江212001 [2]南京医科大学第一附属医院血液科,江苏南京210029 [3]江苏大学药学院,江苏镇江212001

出　　处：《南京医科大学学报（自然科学版）》2012年第2期177-182,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省自然科学基金(BK2008236);镇江市社会发展项目(SH2010030;SH2011021)

摘　　要：目的:研究正常骨髓间充质干细胞(mesenchymal stem cell,MSC)在与骨髓瘤细胞株U266、RPMI8226共培养过程中,骨髓瘤细胞对MSC EphB4/ephrinB2表达的影响。方法:应用Transwell培养体系,将正常骨髓MSC分别与骨髓瘤细胞株U266、RPMI8226细胞共培养7、12 d后分别应用实时定量RT-PCR和细胞免疫化学方法检测骨髓MSC的EphB4/ephrinB2mRNA和蛋白表达水平。结果:骨髓MSC与U266、RPMI8226共培养后,与对照组MSC相比,生长和形态未见明显改变,应用实时定量RT-PCR方法检测骨髓MSC EphB4/ephrinB2的mRNA水平较对照组均降低,在共培养7 d的时间点上除EphB4在RPMI8226共培养组与对照组差异无统计学意义,其余组间差异均有统计学意义(P<0.05),在12 d上差异无统计学意义(P>0.05)。细胞免疫化学结果显示共培养组MSC胞膜胞浆EphB4/ephrinB2表达下降。结论:骨髓瘤细胞株U266、RPMI8226在与正常骨髓MSC共培养后,诱导骨髓MSC EphB4/ephrinB2表达的下调,可能通过MSC成骨-破骨活动的脱耦合,参与骨髓瘤骨病的发生。Objective： To investigate the expression changes of EphB4/ephrinB2 in normal bone marrow derived mesenchymal stem ceils （MSCs） co-cultured with myeLoma cell lines U266 or RPMI8226. Methods： Bone marrow MSCs from normal subjects were co-cultured with U266 or RPMI8226 by transwell for 7 days to 12 days. The mRNA expression levels of EphB4/ephrinB2 in MSCs were detected by real time quantitative PCR（RT-PCR） at point times and the protein levels of EphB4/ephrinB2 were analyzed by immunocytochemistry. Results： No evident morphologic and proliferative alterations could be observed in MSCs after co-culture with U266 or RPMI8226. The mean mRNA expression levels of EphB4/ephrinB2 were down-regulated in MSCs after co-cuLture with U266 or RPMI8226 as compared with controls. There was a significant difference on the point time of the 7th day＇ point（P 〈 0.05） except EphB4 of MSCs co-cultured with RPMI8226 and no significant difference on the 12 day＇ point（P 〉 0.05） between co-cultured MSCs and the controls. The protein Levels of EphB4/ephrinB2 were also reduced in co-cultured MSCs. Conclusion： The communication between bone marrow MSCs and myeloma cells induced dysregulation of EphB4/ephrinB2,which may contribute to uncoupling of bone remodeling in myeloma bone disease.

关 键 词：骨髓瘤骨病 间充质干细胞 EPHB4 EPHRINB2 

分 类 号：Q786[生物学—分子生物学] R738.1[医药卫生—肿瘤]