骨髓间充质干细胞与骨髓瘤细胞共培养时EphB4/ephrinB2表达异常
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江苏省自然科学基金( BK2008236);镇江市社会发展项目(SH2010030,SH2011021)


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

    Abstract:

    Objective: To investigate the expression changes of EphB4/ephrinB2 in normal bone marrow derived mesenchymal stem cells (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.

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王 欢,李 皎,朱 彦,陆 化,童珊珊,余先球,王丽霞,汤 郁,费小明.骨髓间充质干细胞与骨髓瘤细胞共培养时EphB4/ephrinB2表达异常[J].南京医科大学学报(自然科学版),2012,(2):177-182

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  • 收稿日期:2011-10-09
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