Department of Epidemiology and Biostatistics, School of Public Health, Xi'an Jiao Tong University Health Science Center, Xi'an, Shaanxi 710061, China; College of Public Health and Henan Key Laboratory of Tumor Epidemiology, Zhengzhou University, Zhengzhou, Henan 450052, China.
Department of Clinical Laboratory, The First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052, China.
Biochem Biophys Res Commun. 2018 Sep 5;503(2):452-458. doi: 10.1016/j.bbrc.2018.04.094. Epub 2018 Jun 30.
Esophageal cancer (EC) is one of the most common malignancies with high incidence and mortality. Tumor-associated macrophages (TAMs) in the tumor microenvironment have been linked to the accelerated tumor progression. MicroRNAs (miR) are 19-25 nucleotide-long, noncoding RNA molecules, functioning as modulators of gene expression, and mediate a variety of biological functions, including tumor growth. In the present study, the effects and molecular mechanism of miR-155 in TAMs isolated from EC were explored. The expression of miR-155 and fibroblast growth factor-2 (FGF2) in EC tissues and cell lines were analyzed using reverse transcription-quantitative PCR (qRT-PCR) and western blot assays. TAMs were also transfected with the described constructs. Following, the culture medium from TAMs was collected for further analysis. The released FGF2, and inflammatory cytokines were quantified using ELISA. The cell viability, migrated and invaded levels were calculated through Cell Counting kit-8 (CCK8), and transwell analysis. Moreover, human umbilical vein endothelial cells (HUVEC) vasculature formation was determined using matrigel angiogenesis analysis. The results indicated that miR-155 expression was decreased in EC tissues and cell lines, while FGF2 expression was increased in comparison to those in the normal control group. Moreover, miR-155 mimics transfection up-regulated tumor necrosis factor α (TNF-α), interleukin (IL)-12 and inducible nitric oxide synthase (iNOS), while down-regulated IL-10, Arginase-1 (Arg-1) and IL-22 levels in the culture medium from TAMs. And enhancing miR-155 expression in TAMs suppressed the cell viability, migration and invasion of ECA109 cells and reduced the angiogenesis. Nevertheless, over-expressing FGF2 abolished the role of miR-155 in cancer cell survival, migration, invasion as well as angiogenesis. Our findings indicated that miR-155-regulated FGF2 expression from TAMs suppressed EC cell proliferation, migration, invasion and inhibited vasculature formation. Thus, miR-155-modulated FGF2 might be a potential therapeutic target to prevent EC progression.
食管癌（EC）是发病率和死亡率都很高的最常见恶性肿瘤之一。肿瘤微环境中的肿瘤相关巨噬细胞（TAMs）与肿瘤的加速进展有关。微小 RNA（miR）是长约 19-25 个核苷酸的非编码 RNA 分子，作为基因表达的调节剂，调节多种生物学功能，包括肿瘤生长。本研究探讨了 miR-155 在 EC 分离的 TAMs 中的作用及其分子机制。采用逆转录定量 PCR（qRT-PCR）和 Western blot 检测 miR-155 和成纤维细胞生长因子 2（FGF2）在 EC 组织和细胞系中的表达。TAMs 也用所述构建体转染。随后，收集 TAMs 的培养上清液进行进一步分析。采用 ELISA 定量检测释放的 FGF2 和炎性细胞因子。通过细胞计数试剂盒-8（CCK8）和 Transwell 分析计算细胞活力、迁移和侵袭水平。此外，用人脐静脉内皮细胞（HUVEC）血管生成分析确定血管生成。结果表明，与正常对照组相比，miR-155 在 EC 组织和细胞系中的表达降低，而 FGF2 的表达升高。此外，miR-155 模拟物转染上调 TAMs 培养上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-12 和诱导型一氧化氮合酶（iNOS）的表达，下调白细胞介素（IL）-10、精氨酸酶-1（Arg-1）和白细胞介素（IL）-22 的表达。增强 TAMs 中的 miR-155 表达抑制了 ECA109 细胞的活力、迁移和侵袭，并减少了血管生成。然而，过表达 FGF2 消除了 miR-155 在癌细胞存活、迁移、侵袭和血管生成中的作用。我们的研究结果表明，miR-155 调节 TAMs 中的 FGF2 表达，抑制 EC 细胞增殖、迁移、侵袭，并抑制血管生成。因此，miR-155 调节的 FGF2 可能是预防 EC 进展的潜在治疗靶点。
