Silva Marília, Monteiro Gabriel Amaro, Fialho Arsenio M, Bernardes Nuno, da Silva Cláudia Lobato
iBB-Institute for Bioengineering and Biosciences, Department of Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
Front Cell Dev Biol. 2020 Jul 9;8:471. doi: 10.3389/fcell.2020.00471. eCollection 2020.
Recently, cell-based therapies have been explored as a strategy to enhance the specificity of anticancer therapeutic agents. In this perspective, human mesenchymal stromal cells (MSC) hold a promising future as cell delivery systems for anticancer proteins due to their unique biological features. In this study, we engineered human MSC to secrete a human codon-optimized version of azurin (hazu), a bacterial protein that has demonstrated anticancer activity toward different cancer models both and . To this end, microporation was used to deliver plasmid DNA encoding azurin into MSC derived from bone marrow (BM) and umbilical cord matrix (UCM), leading to expression and secretion of hazu to the conditioned medium (CM). Engineered hazu-MSC were shown to preserve tumor tropism toward breast (MCF-7) and lung (A549) cancer cell lines, comparable to non-modified MSC. Azurin was detected in the CM of transfected MSC and, upon treatment with hazu-MSC-CM, we observed a decrease in cancer cell proliferation, migration, and invasion, and an increase in cell death for both cancer cell lines. Moreover, expression of azurin caused no changes in MSC expression profile of cytokines relevant in the context of cancer progression, thus suggesting that the antitumoral effects induced by hazu-MSC secretome might be due to the presence of azurin independently. In conclusion, data shown herein indicate that MSC-produced azurin in a CM configuration elicits an anticancer effect.
最近，基于细胞的疗法已被探索作为一种增强抗癌治疗剂特异性的策略。从这个角度来看，人间充质基质细胞（MSC）因其独特的生物学特性，作为抗癌蛋白的细胞递送系统具有广阔的前景。在本研究中，我们对人MSC进行基因工程改造，使其分泌人密码子优化的天青蛋白（hazu），这是一种细菌蛋白，已在体内和体外对不同癌症模型显示出抗癌活性。为此，采用微孔电穿孔法将编码天青蛋白的质粒DNA导入源自骨髓（BM）和脐带基质（UCM）的MSC，导致hazu在条件培养基（CM）中表达和分泌。工程化的hazu-MSC显示出对乳腺癌（MCF-7）和肺癌（A549）细胞系保持肿瘤嗜性，与未修饰的MSC相当。在转染的MSC的CM中检测到天青蛋白，在用hazu-MSC-CM处理后，我们观察到两种癌细胞系的癌细胞增殖、迁移和侵袭减少，细胞死亡增加。此外，天青蛋白的表达未引起MSC在癌症进展背景下相关细胞因子表达谱的变化，因此表明hazu-MSC分泌组诱导的抗肿瘤作用可能独立于天青蛋白的存在。总之，本文所示数据表明，以CM形式由MSC产生的天青蛋白具有抗癌作用。
