JAMA Ophthalmol. 2014 May;132(5):614-21. doi: 10.1001/jamaophthalmol.2014.43.
Among the characteristics of uveal melanoma that are associated with a poor prognosis are a large tumor size and the presence of increased numbers of lymphocytes and macrophages. In rapidly growing tumors, reduction in oxygen tension may occur with increased distance from blood vessels, which we hypothesize may lead to an inflammatory microenvironment, further stimulating tumor growth.
To analyze whether hypoxia induces uveal melanoma cells to express proinflammatory cytokines and whether tumor supernatant (TSN) affects monocyte migration and differentiation.
The expression of proinflammatory genes in freshly cultured uveal melanoma samples was studied in an in vitro 24-hour hypoxic culture system using quantitative polymerase chain reaction. In addition, cell lines cultured under normoxic and hypoxic conditions were used. The effect of TSN on monocyte chemotaxis was tested using a transwell migration system and by analyzing monocyte differentiation. The levels of the cytokines CCL2, IL6, and PGE2 in TSN were determined by enzyme-linked immunosorbent assay.
Five cell lines (OCM8, 92.1, Mel270,Mel290 and OMM2.5) and 11 primary short-term cultures.
Exposure of freshly cultured uveal melanoma cells to hypoxia led to an increased expression of the proinflammatory cytokines PLGF (OMIM 601121), TGFβ (OMIM 190180), END1 (OMIM +131240), and ICAM1 (OMIM 147840) and a lower expression of AIMP1 (OMIM 603605) (EMAP2), CCL2 (MCP-1) (OMIM +158105), and IL1b (OMIM *147720). The TSN from cultured melanoma cell lines induced chemotaxis of monocytes, but this was independent of the normoxic or hypoxic state. The TSN of 1 cell line and 2 primary uveal melanoma cultures inhibited the dendritic cell maturation and did not induce M2 macrophage polarization in vitro.
Our results indicate that under hypoxic conditions, immune response genes are differentially expressed in cultured primary uveal melanoma cells. The TSN from uveal melanoma cell lines is capable of affecting the chemotactic response and maturation of monocytes in vitro, but this is irrespective of hypoxia.
脉络膜黑色素瘤与预后不良相关的特征之一是肿瘤体积大，以及淋巴细胞和巨噬细胞数量增加。在快速生长的肿瘤中，随着远离血管的距离增加，氧气张力可能会降低，这可能导致炎症微环境，进一步刺激肿瘤生长。
分析缺氧是否会诱导葡萄膜黑色素瘤细胞表达促炎细胞因子，以及肿瘤上清液（TSN）是否会影响单核细胞迁移和分化。
使用定量聚合酶链反应，在体外 24 小时缺氧培养系统中研究新鲜培养的葡萄膜黑色素瘤样本中促炎基因的表达。此外，还使用在常氧和缺氧条件下培养的细胞系。使用 Transwell 迁移系统和分析单核细胞分化来测试 TSN 对单核细胞趋化性的影响。通过酶联免疫吸附试验测定 TSN 中细胞因子 CCL2、IL6 和 PGE2 的水平。
5 个细胞系（OCM8、92.1、Mel270、Mel290 和 OMM2.5）和 11 个短期培养的原发性细胞。
将新鲜培养的葡萄膜黑色素瘤细胞暴露于缺氧环境中，会导致促炎细胞因子 PLGF（OMIM 601121）、TGFβ（OMIM 190180）、END1（OMIM +131240）和 ICAM1（OMIM 147840）的表达增加，而 AIMP1（OMIM 603605）（EMAP2）、CCL2（MCP-1）（OMIM +158105）和 IL1b（OMIM *147720）的表达降低。培养的黑色素瘤细胞系的 TSN 诱导单核细胞趋化，但这与常氧或缺氧状态无关。1 个细胞系和 2 个原发性葡萄膜黑色素瘤培养物的 TSN 抑制树突状细胞成熟，并且不会在体外诱导 M2 巨噬细胞极化。
我们的研究结果表明，在缺氧条件下，培养的原发性葡萄膜黑色素瘤细胞中免疫反应基因的表达存在差异。葡萄膜黑色素瘤细胞系的 TSN 能够影响体外单核细胞的趋化反应和成熟，但这与缺氧无关。
