Abstract：
During cancer evolution, tumor cells attract and dynamically interact with monocytes/macrophages. To find biomarkers of disease progression in human melanoma, we used unbiased RNA sequencing and secretome analyses of tumor-macrophage co-cultures. Pathway analysis of genes differentially modulated in human macrophages exposed to melanoma cells revealed a general upregulation of inflammatory hallmark gene sets, particularly chemokines. A selective group of chemokines, including CCL8, CCL15, and CCL20, was actively secreted upon melanoma-macrophage co-culture. Because we previously described the role of CCL20 in melanoma, we focused our study on CCL8 and CCL15 and confirmed that in vitro both chemokines contributed to melanoma survival, proliferation, and 3D invasion through CCR1 signaling. In vivo, both chemokines enhanced primary tumor growth, spontaneous lung metastasis, and circulating tumor cell survival and lung colonization in mouse xenograft models. Finally, we explored the clinical significance of CCL8 and CCL15 expression in human skin melanoma, screening a collection of 67 primary melanoma samples, using multicolor fluorescence and quantitative image analysis of chemokine-chemokine receptor content at the single-cell level. Primary skin melanomas displayed high CCR1 expression, but there was no difference in its level of expression between metastatic and nonmetastatic cases. By contrast, comparative analysis of these two clinically divergent groups showed a highly significant difference in the cancer cell content of CCL8 (p = 0.025) and CCL15 (p < 0.0001). Kaplan-Meier curves showed that a high content of CCL8 or CCL15 in cancer cells correlated with shorter disease-free and overall survival (log-rank test, p < 0.001). Our results highlight the role of CCL8 and CCL15, which are highly induced by melanoma-macrophage interactions in biologically aggressive primary melanomas and could be clinically applicable biomarkers for patient profiling. © 2024 The Pathological Society of Great Britain and Ireland.
摘要：
在癌症进化过程中,肿瘤细胞吸引并动态地与单核细胞/巨噬细胞相互作用。为了找到人类黑色素瘤疾病进展的生物标志物，我们使用无偏RNA测序和肿瘤-巨噬细胞共培养物的分泌组分析.在暴露于黑色素瘤细胞的人巨噬细胞中差异调节的基因的通路分析揭示了炎症标志基因集的普遍上调。特别是趋化因子。一组选择性的趋化因子，包括CCL8,CCL15和CCL20在黑素瘤-巨噬细胞共培养后被主动分泌.因为我们之前描述了CCL20在黑色素瘤中的作用，我们的研究集中在CCL8和CCL15上，并证实在体外两种趋化因子都有助于黑色素瘤的存活，扩散,和3D入侵通过CCR1信号。在体内,两种趋化因子都能促进原发性肿瘤的生长，自发性肺转移,和小鼠异种移植模型中循环肿瘤细胞存活和肺定植。最后,我们探讨了CCL8和CCL15在人类皮肤黑色素瘤中的表达的临床意义，筛选了67个原发性黑色素瘤样本，使用多色荧光和定量图像分析趋化因子-趋化因子受体在单细胞水平上的含量。原发性皮肤黑素瘤显示高CCR1表达，但其表达水平在转移性和非转移性病例之间没有差异。相比之下,这两个临床差异组的比较分析显示，CCL8（p=0.025）和CCL15（p<0.0001）的癌细胞含量存在高度显着差异。Kaplan-Meier曲线表明，癌细胞中CCL8或CCL15的高含量与较短的无病生存期和总生存期相关（对数秩检验，p<0.001)。我们的结果强调了CCL8和CCL15的作用，它们在生物学侵袭性原发性黑色素瘤中由黑色素瘤-巨噬细胞相互作用高度诱导，并且可能是临床上适用于患者概况的生物标志物。©2024英国和爱尔兰病理学会。
