多发性骨髓瘤患者骨髓间充质干细胞在体外对骨髓瘤细胞趋化迁移的影响

Effect of Bone Marrow Mesenchymal Stem Cells in Patients with Multiple Myelima on Migration of Myeloma Cells In Vitro

目的:探讨体外培养情况下多发性骨髓瘤患者骨髓间充质干细胞(MSC)对骨髓瘤细胞趋化迁移的影响。方法:采用体外培养方法,将骨髓瘤细胞株U266分成两组:A组与正常人骨髓间充质干细胞(N-MSC)共培养,B组与多发性骨髓瘤患者的骨髓间充质干细胞共培养,在有或无硼替佐米作用条件下比较2组U266细胞株的CCR1表达水平、Transwell迁移率以及N-MSC或MM-MSC培养液上清中U266细胞在Transwell中的迁移情况。结果:U266细胞与N-MSC组和MM-MSC共培养后,B组U266细胞在Transwell中的迁移率较高(P<0.05);硼替佐米处理U266细胞后不能消除2组的区别;B组U266细胞的CCR1表达水平高于A组(P<0.05)。骨髓MSC培养上清液实验显示,在无硼替佐米作用条件下,MM-MSC和N-MSC培养上清液与SDF-1相比,具有较强的趋化刺激能力,使细胞在Transwell中的迁移率增高,而在有硼替佐米作用下,3组培养上清液使细胞在Transwell中迁移率降低(P<0.05),但不管是否在硼替佐米作用下,MM-MSC和N-MSC培养上清液对U266细胞在Transw ell中的迁移作用无显著性影响(P>0.05)。结论:骨髓瘤患者的骨髓间充质干细胞本身有部分内在缺陷,通过与骨髓瘤细胞直接相互作用而影响其体外趋化功能。

Objective： To explore the effect of bone marrow mesenchymal stem cells （MSC） in patients with multiple myeloma （MM） on chemotactic migration of myeloma cells in vitro. Methods： By in vitro co-culture with diffferent MSC, the myeloma cell U266 was divided into 2 groups： group A in which the U266 cells were co-cultured with normal person MSC （N-MSC） and group B in which the U266 cells were co-cultured with MM-MSC. The expression level of CCR1 in U266 cells, migration rate of U266 cells in Transwell, and the effect of supernantant from co-culture of U266 cells with N-MSC and MM-MSC on the migration in Transwell were compared in condition with or without bortezomib. Results：After co-culture of U266 ceils with N - MSC or MM - MSC, the migration rate of U266 cells in Transwell in B group was higher than that in A group（ P 〈 0. 05 ）. The difference between 2 groups could not be eliminated after treatment of U266 cells with bortezomib. The CCR1 expression level of U266 cells in B group was higher than that in A group （ P 〈 0. 05 ）. The culture supernatant of bone marrow MSC showed that in condition without bortezomib the culture supernatant of MSC in MM patients and normal persons both possessed more strong chemotactic ability and enhanced the migration rate of ceils in Transwell, compared with SDF-1, meanswhile the culture supernatant in 3 groups reduced the migration rate of cells in condition with bortezomib （ P 〈 0. 05 ）, but there were no statistical difference in migration rate of U266 cells in Transwell between supernatant of N - MSC and MM - MSC culture （ P 〉 0. 05）, no matter the bortezonib was used or not. Conclusion： The bone marrow MSC in MM patients have same intrinsic defects that affect the chemotaxis of cells in vitro by directly interacting with myeloma cells.