Pediatric Surgery Department, Children's Hospital G. Di Cristina, ARNAS Civico-Di Cristina-Benfratelli, Via dei Benedettini n.1, 90134, Palermo, Italy.
Cellular and Molecular Pathophysiology Laboratory, Department of Surgical, Oncological and Stomatological Sciences, University of Palermo, Palermo, Italy.
BMC Cancer. 2018 Nov 27;18(1):1176. doi: 10.1186/s12885-018-5082-2.
It has been proposed that mesenchymal stromal cells (MSCs) promote tumor progression by interacting with tumor cells and other stroma cells in the complex network of the tumor microenvironment. We characterized MSCs isolated and expanded from tumor tissues of pediatric patients diagnosed with neuroblastomas (NB-MSCs) to define interactions with the tumor microenvironment.
Specimens were obtained from 7 pediatric patients diagnosed with neuroblastoma (NB). Morphology, immunophenotype, differentiation capacity, proliferative growth, expression of stemness and neural differentiation markers were evaluated. Moreover, the ability of cells to modulate the immune response, i.e. inhibition of phytohemagglutinin (PHA) activated peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cytotoxic function, was examined. Gene expression profiles, known to be related to tumor cell stemness, Wnt pathway activation, epithelial-mesenchymal transition (EMT) and tumor metastasis were also evaluated. Healthy donor bone marrow-derived MSCs (BM-MSC) were employed as controls.
NB-MSCs presented the typical MSC morphology and phenotype. They showed a proliferative capacity superimposable to BM-MSCs. Stemness marker expression (Sox2, Nanog, Oct3/4) was comparable to BM-MSCs. NB-MSC in vitro osteogenic and chondrogenic differentiation was similar to BM-MSCs, but NB-MSCs lacked adipogenic differentiation capacity. NB-MSCs reached senescence phases at a median passage of P7 (range, P5-P13). NB-MSCs exhibited greater immunosuppressive capacity on activated T lymphocytes at a 1:2 (MSC: PBMC) ratio compared with BM-MSCs (p = 0.018). NK cytotoxic activity was not influenced by co-culture, either with BM-MSCs or NB-MSCs. Flow-cytometry cell cycle analysis showed that NB-MSCs had an increased number of cells in the G0-G1 phase compared to BM-MSCs. Transcriptomic profiling results indicated that NB-MSCs were enriched with EMT genes compared to BM-MSCs.
We characterized the biological features, the immunomodulatory capacity and the gene expression profile of NB-MSCs. The NB-MSC gene expression profile and their functional properties suggest a potential role in promoting tumor escape, invasiveness and metastatic traits of NB cancer cells. A better understanding of the complex mechanisms underlying the interactions between NB cells and NB-derived MSCs should shed new light on potential novel therapeutic approaches.
有人提出，间充质基质细胞（MSCs）通过与肿瘤细胞和肿瘤微环境复杂网络中的其他基质细胞相互作用，促进肿瘤的进展。我们对从诊断为神经母细胞瘤（NB）的儿科患者的肿瘤组织中分离和扩增的 MSCs 进行了特征描述，以确定与肿瘤微环境的相互作用。
从 7 名诊断为神经母细胞瘤（NB）的儿科患者中获得标本。评估了形态、免疫表型、分化能力、增殖生长、干性和神经分化标志物的表达。此外，还检查了细胞调节免疫反应的能力，即抑制植物血球凝集素（PHA）激活的外周血单核细胞（PBMC）和自然杀伤（NK）细胞的细胞毒性功能。还评估了与肿瘤细胞干性、Wnt 通路激活、上皮间质转化（EMT）和肿瘤转移相关的基因表达谱。健康供体骨髓来源的 MSCs（BM-MSC）被用作对照。
NB-MSCs 呈现出典型的 MSC 形态和表型。它们表现出与 BM-MSCs 相似的增殖能力。干性标志物的表达（Sox2、Nanog、Oct3/4）与 BM-MSCs 相当。NB-MSC 的体外成骨和成软骨分化与 BM-MSCs 相似，但 NB-MSCs 缺乏成脂分化能力。NB-MSCs 在中位数为 P7（范围，P5-P13）的传代中达到衰老阶段。与 BM-MSCs 相比，NB-MSCs 在 1:2（MSC：PBMC）比例时对激活的 T 淋巴细胞具有更强的免疫抑制能力（p=0.018）。共培养对 NK 细胞的细胞毒性活性没有影响，无论是与 BM-MSCs 还是 NB-MSCs 共培养。流式细胞术细胞周期分析显示，与 BM-MSCs 相比，NB-MSCs 中处于 G0-G1 期的细胞数量增加。转录组谱分析结果表明，与 BM-MSCs 相比，NB-MSCs 富含 EMT 基因。
我们对 NB-MSCs 的生物学特征、免疫调节能力和基因表达谱进行了描述。NB-MSC 的基因表达谱及其功能特性表明，它们在促进 NB 癌细胞的肿瘤逃逸、侵袭和转移特性方面可能发挥作用。更好地了解 NB 细胞与 NB 来源的 MSC 之间相互作用的复杂机制，应该能为新的潜在治疗方法提供新的认识。
