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首页> 美国卫生研究院文献>Antioxidants >Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions
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Gene and Protein Expression Is Altered by Ascorbate Availability in Murine Macrophages Cultured under Tumour-Like Conditions

机译：通过在肿瘤样条件下培养的鼠巨噬细胞中的抗坏血性可用性来改变基因和蛋白质表达

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Tumour-associated macrophages (TAMs) are ubiquitously present in tumours and commonly associated with poor prognosis. In immune cells, ascorbate affects epigenetic regulation, differentiation and phenotype via its co-factor activity for the 2-oxoglutarate dependent dioxygenase enzymes. Here, we determined the effect of ascorbate on TAM development in response to tumour microenvironmental cues. Naïve murine bone marrow monocytes were cultured with Lewis Lung Carcinoma conditioned media (LLCM) or macrophage colony-stimulating factor (MCSF) to encourage the development into tumour-associated macrophages. Cells were stimulated with hypoxia (1% O2), with or without ascorbate (500 µM) supplementation. Cells and media were harvested for gene, cell surface marker and protein analyses. LLCM supported bone marrow monocyte growth with >90% of cells staining CD11b+F4/80+, indicative of monocytes/macrophages. LLCM-grown cells showed increased expression of M2-like and TAM genes compared to MCSF-grown cells, which further increased with hypoxia. In LLCM-grown cells, ascorbate supplementation was associated with increased F4/80 cell surface expression, and altered gene expression and protein secretion. Our study shows that ascorbate modifies monocyte phenotype when grown under tumour microenvironmental conditions, but this was not clearly associated with either a pro- or anti-tumour phenotype, and reflects a complex and nuanced response of macrophages to ascorbate. Overall, ascorbate supplementation clearly has molecular consequences for TAMs, but functional and clinical consequences remain unknown.
机译：肿瘤相关的巨噬细胞（TAMS）普遍存在于肿瘤中，通常与预后不良相关。在免疫细胞中，抗​​坏血酸物通过其与2-氧氧化术依赖性二氧化酶的共同因子活性影响表观遗传调控，分化和表型。在这里，我们确定抗坏血酸对肿瘤微环境提示的影响对TAM发育的影响。 Naïve鼠骨髓单核细胞用Lewis肺癌条件培养基（LLCM）或巨噬细胞菌落刺激因子（MCSF）培养，以促进肿瘤相关的巨噬细胞的发育。用缺氧（1％O 2）刺激细胞，用或不含抗坏血酸（500μm）补充剂。收获细胞和培养基，用于基因，细胞表面标记和蛋白质分析。 LLCM支持骨髓单核细胞生长，染色CD11b + F4 / 80 +的90％，指示单核细胞/巨噬细胞。与MCSF生长的细胞相比，LLCM生长的细胞显示出类似的M2样和TAM基因的表达，其与缺氧进一步增加。在LLCM生长的细胞中，抗​​坏血酸补充剂与增加的F4 / 80细胞表面表达，并改变基因表达和蛋白质分泌。我们的研究表明，当在肿瘤微环境条件下生长时，抗坏血酸改变单核细胞表型，但这与抗肿瘤表型没有明显相关，反映巨噬细胞对抗坏血酸的复杂性和细致感应者。总体而言，抗坏血酸补充剂显然对TAMS进行了分子后果，但功能性和临床后果仍然未知。

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