[Effect of γδ T cells on the Proliferation, Apoptosis and Autophagy of Multiple Myeloma Cells].

AbstractObjective: To investigate the effect of γδ T cells on the proliferation, apoptosis and autophagy of multiple myeloma cells.Peripheral blood mononuclear cells (PBMNC) were isolated from healthy volunteers, and stimulated with zoledronic acid (Zol) in combination with rhIL-2. Flow cytometry analysis was used to detected the purity of γδ T cells. γδ T cells were collected and co-cultured with RPMI-8226 or U-266 cells at different effector target ratios. The proliferation of RPMI-8226 or U-266 cell lines were detected by CCK-8. Cell cycle and cell apoptosis were detected by flow cytometry and Western blot．The expressions of autophagy-related proteins were detected by Western blot.γδ T cells can be expanded in vitro. γδ T cells could inhibit the proliferation of RPMI-8226 or U-266 cells, induced cell cycle arrest and promoted apoptosis in an effector target-dependent manner. In addition, γδ T cells could induce autophagy of myeloma cells, inhibited the expression of autophagy-related PI3K, P-AKT and P-mTOR, while increased the expression of AMPK and Beclin-1.γδ T cells can inhibit the proliferation of RPMI-8226 and U-266 myeloma cells, induce cell cycle arrest, promote apoptosis, and enhance autophagy in vitro. The mechanism may be related to inhibition of PI3K/AKT/mTOR signaling pathway and/or activation of AMPK/Beclin-1 signaling pathway.γδ T细胞调控多发性骨髓瘤细胞增殖、凋亡和自噬的作用.探讨γδ T细胞对多发性骨髓瘤细胞增殖、凋亡及自噬的影响.采用Ficoll分离法获取健康成年人外周血单个核细胞（PBMNC），体外刺激诱导获取γδ T细胞，流式细胞术检测γδ T细胞纯度。收集γδ T细胞与骨髓瘤细胞RPMI-8226或U-266按照不同效靶比于transwell小室中分别共培养。采用CCK-8法检测细胞增殖，通过流式细胞术和Western blot方法检测细胞周期分布和细胞凋亡情况，通过Western blot方法检测自噬相关蛋白表达情况.PBMNC在体外刺激诱导下，可获得大量扩增的γδ T细胞。γδ T细胞具有抑制骨髓瘤细胞增殖的作用，同时诱导细胞周期阻滞并促进细胞的凋亡，且作用具有效靶比依赖性。此外，γδ T细胞可增强骨髓瘤细胞的自噬能力，抑制骨髓瘤细胞PI3K、P-AKT和P-mTOR的表达，上调AMPK和Beclin-1的表达水平.γδ T细胞在体外具有抑制骨髓瘤细胞RPMI-8226或U-266的增殖，促进其凋亡和增强自噬的作用。其作用机制可能是通过抑制PI3K/AKT/mTOR信号通路，和/或激活AMPK/Beclin-1信号通路.