TNF-α derived from M2 tumor-associated macrophages promotes epithelial-mesenchymal transition and cancer sternness through the Wnt/β-catenin pathway in SMMC-7721 hepatocellular carcinoma cells
TNF-α derived from M2 tumor-associated macrophages promotes epithelial-mesenchymal transition and cancer sternness through the Wnt/β-catenin pathway in SMMC-7721 hepatocellular carcinoma cells
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源自 M2 肿瘤相关巨噬细胞的 TNF-α 通过 Wnt/β-catenin 通路促进 SMMC-7721 肝癌细胞上皮间质转化和癌症恶化
DOI：
10.1016/j.yexcr.2019.03.005
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发表时间：
2019-05-01
影响因子：
3.7
通讯作者：
Wang, Changjun
中科院分区：
文献类型：
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作者：
Chen, Yongxu;Wen, Huihong;Wang, Changjun
M2极化的肿瘤相关巨噬细胞（M2-TAM）浸润肿瘤微环境有助于肝细胞癌（HCC）的进展。据报道，经历EMT的癌细胞将获得干性特征。在此，将HCC SMMC-7721细胞系与从THP-1细胞极化的M2-TAM在体外共培养。在体内研究中，我们使用裸鼠皮下肿瘤模型来测试肿瘤的生长是否受到M2-TAMs的影响。随后，在细胞和皮下肿瘤组织中检测EMT、干细胞和Wnt/β-连环蛋白通路相关标志物。还在共培养系统上清液和裸鼠血清中评估TNF-α。我们发现SMMC-7721在与M2-TAMs共培养后经历EMT并获得干性，并且与单独的SMMC-7721相比，皮下注射悬浮在M2-TAMs上清液中的SMMC-7721后导致更大的肿瘤尺寸。酶联免疫吸附试验显示，TNF-α在M2-TAMs上清液中的表达升高，并与裸鼠血清中肿瘤的大小呈正相关。此外，我们发现Wnt/β-连环蛋白途径是TNF-α的下游靶点，并且Wnt/β-连环蛋白抑制剂ICG-001部分逆转EMT并减弱癌症干性。我们的研究结果表明，来自M2-TAMs的TNF-α通过Wnt/β-连环蛋白途径促进EMT和癌症干细胞。
M2-polarized tumor-associated macrophages (M2-TAMs) infiltrating the tumor microenvironment contribute to hepatocellular carcinoma (HCC) progression. It was reported that cancer cells undergoing EMT will acquire sternness characteristics. Here, the HCC SMMC-7721 cell line was co-cultured with M2-TAMs polarized from THP-1 cells in vitro. In in vivo studies, we used nude mice subcutaneous tumor model to test whether the growth of the tumor was affected by M2-TAMs. Subsequently, EMT, sternness and Wnt/beta-catenin pathway related markers were detected in cells and subcutaneous tumor tissues. TNF-alpha was also assessed in both the co-culture system supernatants and in nude mice serum. We found that SMMC-7721 underwent EMT and acquired sternness after co-culture with M2-TAMs, and resulted in larger tumor size following subcutaneous injection of SMMC-7721 suspended in M2-TAMs supernatants compared with SMMC-7721 alone. Enzyme linked immunosorbent assay showed that TNF-alpha expression was elevated in supernatants of M2-TAMs and positively correlated with tumor size in the serum of nude mice. Furthermore, we found that the Wnt/beta-catenin pathway was a downstream target of TNF-alpha and that the Wnt/beta-catenin inhibitor ICG-001 partially reversed EMT and attenuated cancer sternness. Our results indicate that TNF-alpha derived from M2-TAMs promote EMT and cancer sternness cells via the Wnt/beta-catenin pathway.