Department of Hepatobiliary Surgery, Shandong Cancer Hospital Affiliated to Shandong University, Shandong Academy of Medical Sciences, Jinan 250117, Shandong, China.
Department of Hepatobiliary and Pancreatic Surgery, Xuzhou Cancer Hospital, Xuzhou 221000, Jiangsu, China.
Biochimie. 2018 Jul;150:23-30. doi: 10.1016/j.biochi.2018.04.016. Epub 2018 Apr 24.
We aim to investigate the role of miR-98-mediated macrophage polarization in hepatocellular carcinoma (HCC) progression and to explore the underlying mechanism. A total of 25 paired HCC and matched adjacent normal tissues (ANTs) were collected. We incubated human blood monocytes isolated from healthy male donors with culture medium collected from HepG2 cells for 7 days. The mRNA and protein expression was detected by qRT-PCR and Western blot, respectively. Levels of cytokines secreted in culture medium were measured using the specific ELISA kits. The miR-98 mimic/inhibitor was transfected to explore the role of miR-98 in HCC-conditioned tumor-associated macrophages (TAMs). HepG2 cells were then cultured with condition medium from HCC-conditioned TAMs pretreated with miR-98 mimic/inhibitor, and cell migration and invasion assays were performed. Luciferase reporter assay was performed to analyze the interaction between miR-98 and interleukin (IL)-10. Our results showed that miR-98 was downregulated and IL-10 was upregulated in HCC tissues and HCC-conditioned TAMs. Further studies identified that IL-10 was a direct target gene of miR-98 in HCC-conditioned TAMs. Moreover, miR-98 regulated the levels of inflammatory cytokines in HCC-conditioned TAMs. HCC-conditioned TAMs pretreated with miR-98 regulated migration and invasion of HepG2 cells in vitro, and the effects were significantly reversed by IL-10. In conclusion, miR-98 not only regulated expression of inflammatory cytokines in HCC-conditioned TAMs, but also modulated the capacity of HCC-conditioned TAMs to regulate HepG2 cell migration and invasion, at least in part, by targeting IL-10. As a result, miR-98 may suppress the progress of HCC.
我们旨在研究 miR-98 介导的巨噬细胞极化在肝细胞癌 (HCC) 进展中的作用，并探讨其潜在机制。共收集了 25 对 HCC 及其匹配的相邻正常组织 (ANTs)。我们将从健康男性供体中分离出的人血单核细胞与 HepG2 细胞培养的培养基共培养 7 天。通过 qRT-PCR 和 Western blot 分别检测 mRNA 和蛋白质的表达。使用特定的 ELISA 试剂盒测量培养基中细胞因子的分泌水平。转染 miR-98 模拟物/抑制剂以探索 miR-98 在 HCC 条件化肿瘤相关巨噬细胞 (TAMs) 中的作用。然后将 HepG2 细胞与用 miR-98 模拟物/抑制剂预处理的 HCC 条件化 TAMs 的条件培养基一起培养，并进行细胞迁移和侵袭实验。进行荧光素酶报告实验分析 miR-98 与白细胞介素 (IL)-10 之间的相互作用。我们的结果表明，miR-98 在 HCC 组织和 HCC 条件化 TAMs 中下调，而 IL-10 上调。进一步的研究表明，IL-10 是 HCC 条件化 TAMs 中 miR-98 的直接靶基因。此外，miR-98 调节 HCC 条件化 TAMs 中炎症细胞因子的水平。用 miR-98 预处理的 HCC 条件化 TAMs 体外调节 HepG2 细胞的迁移和侵袭，而 IL-10 显著逆转了这些作用。总之，miR-98 不仅调节 HCC 条件化 TAMs 中炎症细胞因子的表达，而且通过靶向 IL-10 调节 HCC 条件化 TAMs 调节 HepG2 细胞迁移和侵袭的能力，至少部分如此。因此，miR-98 可能抑制 HCC 的进展。
