Data_Sheet_1_Conditions Mimicking the Cancer Microenvironment Modulate the Functional Outcome of Human Chorionic Villus Mesenchymal Stem/Stromal Cells in vitro.docx

Mesenchymal stem/stromal cells isolated from chorionic villi of human term placentae (CV-MSCs) possess unique biological characters. They exhibit self-renewal, directional migration, differentiation, and immunomodulatory effects on other cell lineages, by virtue of which they can be utilized as therapeutic carriers, for drug targeting, and therapy. Tumors display characteristic features of a damaged tissue microenvironment, which is saturated with conditions such as hypoxia, sustained inflammation, and increased oxidative stress. CV-MSCs function normally in a high oxidative stress environment induced by hydrogen peroxide (H2O2) and glucose and also protect endothelial cells from their damaging effects. For their therapeutic applications in a disease like cancer, it is necessary to ascertain the effects of tumor microenvironment on their functional outcome. In this study, we investigated the functional activities, of CV-MSCs in response to conditioned media (CM) obtained from the culture of breast cancer cell line MDA-231 (CM-MDA231). CV-MSCs were exposed to CM-MDA231 for different spatio-temporal conditions, and their biological functions as well as modulation in gene expression were evaluated. Effect of CM-MDA231 on factors responsible for changes in functional outcome were also investigated at the protein levels. CV-MSCs exhibited significant reduction in proliferation but increased adhesion and migration after CM-MDA231 treatment. Interestingly, there was no change in their invasion potential. CM-MDA231 treatment modulated expression of various genes involved in important cellular events including, integration, survival, message delivery and favorable outcome after transplantation. Analysis of pathways related to cell cycle regulation revealed significant changes in the expression of p53, and increased phosphorylation of Retinoblastoma (Rb) and Checkpoint Kinase 2 in CV-MSCs treated with CM-MDA231. To summarize, these data reveal that CV-MSCs retain the ability to survive, adhere, and migrate after sustained treatment with CM-MDA231, a medium that mimics the cancer microenvironment. These properties of CV-MSCs to withstand the inflammatory tumor like microenvironment prove that they may make useful candidate in a stem cell based therapy against cancer. However, further pre-clinical studies are needed to validate their therapeutic usage.

从人足月胎盘绒毛膜绒毛分离的间充质干细胞/基质细胞（CV-MSCs）具有独特的生物学特性。它们具备自我更新、定向迁移、分化及对其他细胞谱系的免疫调节作用，凭借这些特性，CV-MSCs可作为治疗载体用于药物靶向与疾病治疗。
肿瘤呈现出受损组织微环境的特征性表现，该微环境处于缺氧、持续炎症及氧化应激增强的状态。CV-MSCs在由过氧化氢（hydrogen peroxide，H₂O₂）和葡萄糖诱导的高氧化应激环境中可维持正常功能，同时能够保护内皮细胞免受氧化应激损伤。鉴于其在癌症等疾病中的治疗应用前景，明确肿瘤微环境对其功能结局的影响十分必要。
本研究中，我们探究了CV-MSCs在响应乳腺癌细胞系MDA-231培养所得条件培养基（conditioned media，CM）刺激后的功能活性。我们将CV-MSCs置于不同时空条件下的CM-MDA231环境中培养，并对其生物学功能及基因表达调控情况进行了评估，同时在蛋白层面探究了CM-MDA231对功能结局相关调控因子的影响。
经CM-MDA231处理后，CV-MSCs的增殖能力显著降低，但黏附与迁移能力有所增强；有趣的是，其侵袭潜能未发生明显变化。CM-MDA231处理可调控多种参与重要细胞事件的基因表达，包括细胞整合、存活、信号传递及移植后良好结局相关的基因。对细胞周期调控相关通路的分析表明，经CM-MDA231处理的CV-MSCs中，p53的表达发生显著改变，视网膜母细胞瘤蛋白（Retinoblastoma，Rb）与检查点激酶2（Checkpoint Kinase 2）的磷酸化水平升高。
综上，本研究数据证实，在持续暴露于模拟癌症微环境的CM-MDA231培养基后，CV-MSCs仍可维持存活、黏附及迁移能力。CV-MSCs能够耐受类似肿瘤的炎性微环境，这一特性表明其有望成为癌症干细胞靶向治疗的潜在候选细胞。不过，仍需开展进一步的临床前研究以验证其治疗应用价值。