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目的研究人多发性骨髓瘤(MM)细胞对内皮细胞分化为管状结构的影响及调控脑源性神经营养因子(BDNF)分泌的初步机制。方法采用人 MM 细胞系 RPMI8226细胞或原代 MM 细胞与人脐静脉内皮细胞(HUVEC)混合共培养和隔离共培养;以同期单独培养的 HUVEC 为对照,对与MM 细胞共培养的 HUVEC 进行 Matrigel 基质管状结构形成实验,评价 MM 细胞对 HUVEC 血管新生能力的影响;采用 ELISA 方法检测两种共培养体系培养上清中 BDNF 的含量;用抗人 CD29和 CD18的单克隆抗体(单抗)对 MM 细胞进行预处理后,在混合共培养的基础上进行管状结构形成实验并检测培养上清中 BDNF 的含量。结果与同期单独培养的 HUVEC 相比,与 RPMI8226细胞隔离共培养的HUVEC 形成的管状结构数量明显增多,但增加幅度(约75%)低于混合共培养体系(约113%)。原代MM 细胞促 HUVEC 管状结构形成效应在隔离共培养体系和混合共培养体系分别为138%与188%。HUVEC 单独培养上清中 BDNF 的含量为(12.4±5.1)ng/ml,RPMI8226细胞混合共培养上清和隔离共培养上清的 BDNF 含量分别为(38.5±8.2)ng/ml 和(31.6±7.2)ng/ml;原代 MM 细胞混合共培养上清和隔离共培养上清的 BDNF 含量分别为(37.1±8.7)ng/ml 和(27.9±7.6)ng/ml。两种黏附分子抗体能不同程度地阻断混合共培养体系中 MM 细胞促 HUVEC 管状结构形成效应,并抑制 BDNF 的分泌。结论 MM 细胞可刺激共培养 HUVEC 分化为管状结构;并受到 MM 细胞与 HUVEC 间可溶性细胞因子和黏附作用的共同调控,其调控机制与 BDNF 相关。
Objective To study the effects of multiple myeloma (MM) cells on the differentiation of endothelial cells into tubular structures and the primary mechanism of regulating brain-derived neurotrophic factor (BDNF) secretion. Methods Human MM cell line RPMI8226 cells or primary MM cells were co-cultured and isolated from human umbilical vein endothelial cells (HUVECs) co-cultured. HUVECs co-cultured with MM cells were treated with Matrigel matrix The effect of MM cells on the angiogenesis ability of HUVECs was evaluated by the formation of tubular structure. The content of BDNF in the culture supernatants of the two co-culture systems was detected by ELISA. The effect of MM cells After pretreatment, the tubular structure formation experiment was conducted on the basis of mixed co-culture and the content of BDNF in the culture supernatant was measured. Results Compared with HUVEC cultured in the same period, the number of tubular structures formed by HUVEC isolated from RPMI8226 cells was significantly increased (about 75%) compared with the mixed co-culture system (about 113%). Primary MM cells promote the formation of HUVEC tubular structure in isolated co-culture system and mixed co-culture system were 138% and 188%. The content of BDNF in the supernatant of HUVEC cultured alone was (12.4 ± 5.1) ng / ml. The contents of BDNF in the mixed culture and isolated co-culture supernatant of RPMI8226 cells were (38.5 ± 8.2) ng / ml and (31.6 ± 7.2) ng / ml. The BDNF contents of primary MM cells mixed co-culture supernatant and isolated co-culture supernatant were (37.1 ± 8.7) ng / ml and (27.9 ± 7.6) ng / ml, respectively. The two adhesion molecule antibodies could block the formation of HUVECs induced by MM cells in mixed co-culture system to a certain extent and inhibit the secretion of BDNF. Conclusions MM cells can stimulate HUVECs to differentiate into tubular structures and be regulated by soluble cytokines and adhesion between MM cells and HUVECs. The regulatory mechanism is related to BDNF.