我们研究了神经母细胞瘤SH-SY5Y细胞的形态和细胞因子谱的特征,骨髓间充质基质/干细胞(BM-MSCs),以及在塑料和基质胶上孵育的双重(BM-MSC+SH-SY5Y细胞)和三重(BM-MSC+SH-SY5Y细胞+PBMC)共培养的外周血单核细胞(PBMC)。共培养物中的细胞通过囊泡运输以及通过交换膜和细胞质成分来传递。在Matrigel和塑料上孵育的双重和三重共培养物的细胞因子谱具有差异,并且显示出多种趋化因子/细胞因子的最高浓度,如CXCL8/IL-8、I-TAC/CXCL11、IP10/CXCL10、MDC/CCL22、MIP-1α/CCL3、IL-1β、ENA-78/CXCL5、Gro-α/CXCL1、MCP-1/CCL2、TERC/CCL25、CXCL8/IL-8和IL-6。三重共培养条件培养基中高浓度的炎症趋化因子/细胞因子形成慢性炎症,这使得提出的共培养系统更接近天然肿瘤。与SH-SY5Y的双重和单一培养相比,三重共培养对顺铂(CDDP)的抗性更高。在双重和三重共培养中共培养和CDDP处理后,细胞中BCL2,BCL2L1,RAC1,CAV1,CASP3和BAX基因的mRNA水平发生变化。BCL2、BAX、测定三重共培养后SH-SY5Y细胞中的CAV1和CASP3蛋白以及双重共培养后SH-SY5Y细胞中的CAV1和BAX蛋白表达。这项研究证明了在我们的肿瘤模型中观察到的肿瘤生态位组分之间的细胞相互作用和细胞间对化学抗性的影响的性质。这应该能够开发用于抗肿瘤剂的新型测试系统。
We investigated the features of the morphology and cytokine profiles of neuroblastoma SH-SY5Y cells, bone marrow-derived mesenchymal stromal/stem cells (BM-MSCs), and peripheral blood mononuclear cells (PBMCs) in double (BM-MSCs + SH-SY5Y cells) and triple (BM-MSCs + SH-SY5Y cells + PBMCs) co-cultures incubated on plastic and Matrigel. Cells in the co-cultures communicated by vesicular transport and by exchanging membrane and cytoplasmic components. The cytokine profile of double and triple co-cultures incubated on Matrigel and plastic had differences and showed the highest concentration of a number of chemokines/cytokines, such as CXCL8/IL-8, I-TAC/CXCL11, IP10/CXCL10, MDC/CCL22, MIP-1α/CCL3, IL-1β, ENA-78/CXCL5, Gro-α/CXCL1, MCP-1/CCL2, TERC/CCL25, CXCL8/IL-8, and IL-6. High concentrations of inflammatory chemokines/cytokines in the conditioned medium of triple co-culture form a chronic inflammation, which brings the presented co-cultivation system closer to a natural tumor. Triple co-cultures were more resistant to cisplatin (CDDP) than the double- and monoculture of SH-SY5Y. The mRNA levels of BCL2, BCL2L1, RAC1, CAV1, CASP3, and BAX genes were changed in cells after co-culturing and CDDP treatment in double and triple co-cultures. The expression of the BCL2, BAX, CAV1, and CASP3 proteins in SH-SY5Y cells after the triple co-culture and CAV1 and BAX protein expression in SH-SY5Y cells after the double co-culture were determined. This study demonstrated the nature of the cellular interactions between components of tumor niche and the intercellular influence on chemoresistance observed in our tumor model, which should enable the development of novel test systems for anti-tumor agents.