Cx43表达与多发性骨髓瘤细胞耐药的相关性

Relationship between connexin-43 expression and drug resistance of multiple myeloma

目的探讨骨髓间充质干细胞(BMSCs)连接蛋白43(Cx43)表达对多发性骨髓瘤(MM)RPMI 8226细胞耐药的影响。方法体外分离、培养BMSCs,采用RT-PCR法检测BMSCs、RPMI 8226细胞Cx43的表达,流式细胞术和多重液相蛋白定量技术分别检测BMSCs与RPMI 8226细胞共培养对RPMI 8226细胞凋亡和细胞因子分泌的影响。结果 BMSCs及RPMI 8226细胞均表达Cx43。BMSCs与RPMI 8226细胞共培养后,上清液中IL-6、IL-10和TGF-β水平增加,硼替佐咪诱导的RPMI 8226细胞凋亡作用减弱;而细胞间隙连接通讯(GJIC)特异性阻断剂18α甘草次酸(18α-GA)能明显逆转上述观察指标的改变过程。结论 BMSCs与RPMI 8226细胞间可形成功能性GJIC,是BMSCs促进MM细胞生存及介导其耐药的重要原因之一。

Objective To investigate the effect of connexin-43（Cx43）expression in bone marrow mesenchymal stem cells（BMSCs）on the drug resistance of multiple myeloma（MM）cell line RPMI 8226.Methods BMSCs were separated and cultured in vitro.Cx43 expression in BMSCs and RPMI 8226 cells was determined by RT-PCR.The effects of BMSCs co-cultured with RPMI 8226 cells on the apoptosis of RPMI 8226 cells and secretion of cytokines were detected by flow cytometry and cytometric beads array,respectively.Results Cx43 was expressed in both BMSCs and RPMI 8226 cells.After BMSCs were co-cultured with RPMI 8226 cells,the levels of IL-6,IL-10 and TGF-βwere increased,while bortezomib-induced apoptosis of RPMI 8226 cells was decreased,which were significantly reversed by gap junction intercellular communication（GJIC）specific inhibitor 18α-glycyrrhetinic acid（18α-GA）.Conclusion The functional GJIC is formed between BMSCs and RPMI 8226 cells,which plays an important role in the survival and drug resistance of MM cells promoted by BMSCs.