Yu Keke, Yang Jinlian, Wang Fei, Chen Li, Lu Yanmin, Luo Jia, Wang Siying
Department of Biaobank, Shanghai Chest Hospital, Shanghai JiaoTong University, Shanghai, China; Department of Pathophysiology, Anhui Medical University, Hefei, Anhui, China; Department of Immunology, Anhui Medical University, Hefei, Anhui, China.
Alcohol Clin Exp Res. 2014 Oct;38(10):2597-606. doi: 10.1111/acer.12512.
Inflammation plays a critical role in cancer progression, and our data suggested that ethanol (EtOH) could promote the progression of breast cancer via increased monocyte chemo-attractant protein-1 (MCP-1). Thus, we investigated the effects of EtOH on lung cancer growth and metastasis to explore whether immunosuppression had a role.
C57BL/6 mice (n = 10) implanted with Lewis lung cancer (LLC) cells were used to model physiologically relevant EtOH intake on tumor growth and inflammation after macrophage polarization. Tumors were isolated and measured, and MCP-1 expression was measured via immunohistochemistry and Western blot. Recruitment of macrophages using CD11b and F4/80 antibodies was detected with immunohistochemistry and flow cytometry. Changes in arginase I and inducible nitric oxide synthase (iNOS) expression were measured with immunofluorescent microscopy. EtOH's effect on in vitro tumor angiogenesis was evaluated in culture, and the tumor microvessel density was assessed with CD31 immunohistochemistry. Matrix metalloproteinase 9 and interleukin 10 expressions were measured by Western blot, ELISA, and immunohistochemistry. Finally, we treated a macrophage cell line RAW264.7 with EtOH and measured changes in arginase I and iNOS expression.
With EtOH exposure, macrophage density was positively correlated with MCP-1 expression. Macrophages infiltrated the tumor site, becoming tumor-associated macrophages that polarized to M2 phenotypes (ArgI(high) /iNOS(low) ) after EtOH treatment. Cancerous cells interacted with immune cells, especially M2 macrophages, and promoted tumor angiogenesis, progression, and invasiveness. RAW264.7 cells stimulated with EtOH expressed more arginase I and less iNOS than controls. These results agreed with the features of M2 phenotype macrophages (ArgI(high) /iNOS(low) ).
Data show that EtOH induced M2 phenotype macrophages, suggesting that progression and metastasis of LLC may be mediated by recruitment of M2 macrophages, especially under the influence of EtOH.
炎症在癌症进展中起关键作用，我们的数据表明乙醇（EtOH）可通过增加单核细胞趋化蛋白-1（MCP-1）促进乳腺癌进展。因此，我们研究了EtOH对肺癌生长和转移的影响，以探讨免疫抑制是否起作用。
将接种Lewis肺癌（LLC）细胞的C57BL/6小鼠（n = 10）用于模拟生理相关的EtOH摄入对巨噬细胞极化后肿瘤生长和炎症的影响。分离并测量肿瘤，通过免疫组织化学和蛋白质印迹法测量MCP-1表达。使用免疫组织化学和流式细胞术检测用CD11b和F4/80抗体招募巨噬细胞的情况。用免疫荧光显微镜测量精氨酸酶I和诱导型一氧化氮合酶（iNOS）表达的变化。在培养中评估EtOH对体外肿瘤血管生成的影响，并用CD31免疫组织化学评估肿瘤微血管密度。通过蛋白质印迹法、酶联免疫吸附测定法和免疫组织化学测量基质金属蛋白酶9和白细胞介素10的表达。最后，我们用EtOH处理巨噬细胞系RAW264.7并测量精氨酸酶I和iNOS表达的变化。
暴露于EtOH后，巨噬细胞密度与MCP-1表达呈正相关。巨噬细胞浸润肿瘤部位，成为肿瘤相关巨噬细胞，在EtOH处理后极化为M2表型（ArgI（高）/iNOS（低））。癌细胞与免疫细胞相互作用，尤其是与M2巨噬细胞相互作用，促进肿瘤血管生成、进展和侵袭。用EtOH刺激的RAW264.7细胞比对照表达更多的精氨酸酶I和更少的iNOS。这些结果与M2表型巨噬细胞（ArgI（高）/iNOS（低））的特征一致。
数据表明EtOH诱导M2表型巨噬细胞，提示LLC的进展和转移可能由M2巨噬细胞的募集介导，尤其是在EtOH的影响下。
