MD2 blockage prevents the migration and invasion of hepatocellular carcinoma cells via inhibition of the EGFR signaling pathway
MD2 blockage prevents the migration and invasion of hepatocellular carcinoma cells via inhibition of the EGFR signaling pathway
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DOI：
10.21037/jgo-21-362
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发表时间：
2021-07-29
影响因子：
2.1
通讯作者：
Fang, Luo
中科院分区：
文献类型：
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作者：
Qi, Yajun;Fang, Qilu;Fang, Luo
背景：Toll样受体（TLR）是肿瘤侵袭和转移中新兴的信号通路。 TLR 的激活需要特定的辅助蛋白，例如小分泌糖蛋白骨髓分化蛋白 2 (MD2)，它有助于配体反应。然而，MD2在肿瘤发生和转移中的作用却鲜有报道。本研究旨在探讨MD2对肝细胞癌（HCC）增殖、迁移和侵袭的影响及其潜在机制。方法：进行细胞计数试剂盒 8 (CCK8)、细胞集落形成、伤口愈合和 Transwell 测定，分别测定细胞活力、增殖、迁移和侵袭。采用实时定量 PCR (qRT-PCR) 评估 HCC 细胞系和人正常肝细胞系中 MD2 的表达以及 MD2 阻断的沉默效率。采用Western blot和qRT-PCR检测上皮间质转化（EMT）标志物和表皮生长因子受体（EGFR）信号分子的蛋白和mRNA表达水平。结果：MD2在HCC组织和细胞系中高表达。 MD2的高表达与HCC患者的不良预后相关。此外，MD2沉默轻微抑制HepG2和HCCLM3的增殖，并显着抑制细胞迁移和侵袭。此外，MD2阻断可以通过增加E-钙粘蛋白和Occludin的蛋白质和mRNA水平，并降低波形蛋白、N-钙粘蛋白和Snail的水平来明显阻止EMT过程。最后，MD2 沉默的 HCC 细胞中 EGFR 及其下游分子 Src、Akt、I-Kappa B α 和 p65 的磷酸化水平下调。结论：我们的研究结果表明，MD2 的高表达可能通过调节 HCC 细胞中的 EGFR 通路来影响 EMT、迁移和侵袭。
Background: The toll-like receptor (TLR) is an emerging signaling pathway in tumor invasion and metastasis. The activation of TLRs requires specific accessory proteins, such as the small secreted glycoprotein myeloid differentiation protein 2 (MD2), which contributes to ligand responsiveness. However, the role of MD2 in tumorigenesis and metastasis has rarely been reported. This study aimed to investigate the effects and underlying mechanisms of MD2 on the proliferation, migration, and invasion of hepatocellular carcinoma (HCC). Methods: Cell counting kit 8 (CCK8), cell colony formation, wound healing, and transwell assays were conducted to determine cell viability, proliferation, migration, and invasion, respectively. Quantitative realtime PCR (qRT-PCR) was performed to assess the expression of MD2 in HCC cell lines and human normal liver cell lines as well as the silencing efficiency of MD2 blockage. Western blot and qRT-PCR assays were performed to detect the protein and mRNA expression levels of epithelial mesenchymal transformation (EMT) markers and epidermal growth factor receptor (EGFR) signaling molecules. Results: MD2 was highly expressed in HCC tissues and cell lines. High expression of MD2 was associated with poor prognosis of HCC patients. In addition, MD2 silencing slightly inhibited the proliferation of HepG2 and HCCLM3, and significantly suppressed cell migration and invasion. Furthermore, MD2 blockage could distinctly prevent the EMT process by increasing the protein and mRNA levels of E-cadherin and Occludin, and decreasing the levels of Vimentin, N-cadherin, and Snail. Finally, the phosphorylation level of EGFR as well as its downstream molecular Src, Akt, I-Kappa B alpha, and p65 were downregulated in HCC cells with MD2 silencing. Conclusions: Our findings suggest that high expression of MD2 may affect the EMT, migration, and invasion via modulation of the EGFR pathway in HCC cells.