ABSTRACT

Aim

In this study, we explored the ability of TAMs to affect the malignant phenotype of human hepatoma Huh-7 cells through the Gli2/IGF-II/ERK1/2 pathway.

Methods

The TAMs were characterized by flow cytometry and ELISA assays. Huh-7 cells were treated with conditioned medium of TAMs (TAMs-CM), and the proliferation, migration and invasion abilities were measured by CCK-8, Transwell and scratch assays. The levels of TGF-β1, Gli2, IGF-II and related proteins in the ERK1/2 pathway and the epithelial-mesenchymal transition (EMT) process were examined by RT-qPCR and western blot. Huh-7 cells were injected subcutaneously into nude mice with TAMs to explore the role of TAMs in tumor growth.

Results

The expression levels of TGF-β1, Gli2 and IGF-II and the cell proliferation, migration and invasion abilities were elevated in Huh-7 cells treated with TAMs-CM. TGF-β1 was upregulated in the conditioned medium and was found to be involved in the promotion of migration, invasion and the EMT of Huh-7 cells. The activation of TGF-β1 signaling increased the expression of Gli2. Knockdown of Gli2 decreased the expression of IGF-II and also reversed the promotional effect of the conditioned medium on migration, invasion and the EMT of Huh-7 cells. TGF-β1/Gli2/IGF-II signaling was shown to promote the malignant phenotype of Huh-7 cells by activating the ERK1/2 signaling pathway. Further, TGF-β1 knockdown attenuated the influence of TAMs on tumor growth in mouse model.

Conclusion

The TGF-β1 secreted by TAMs promotes the migration, invasion and EMT of human hepatoma Huh-7 cells through the Gli2/IGF-II/ERK1/2 pathway.

中文翻译：

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肿瘤相关巨噬细胞通过分泌 TGF-β1 通过 Gli2/IGF-II/ERK1/2 轴促进人肝癌 Huh-7 细胞迁移和侵袭

摘要

目的

在本研究中，我们探索了 TAMs 通过 Gli2/IGF-II/ERK1/2 通路影响人肝癌 Huh-7 细胞恶性表型的能力。

方法

通过流式细胞术和 ELISA 测定对 TAM 进行表征。Huh-7细胞用TAMs条件培养基（TAMs-CM）处理，通过CCK-8、Transwell和scratch法测定其增殖、迁移和侵袭能力。通过RT-qPCR和蛋白质印迹检查ERK1 / 2途径和上皮 - 间质转化（EMT）过程中TGF-β1，Gli2，IGF-II和相关蛋白质的水平。将 Huh-7 细胞皮下注射到含有 TAM 的裸鼠体内，以探索 TAM 在肿瘤生长中的作用。

结果

经TAMs-CM处理的Huh-7细胞中TGF-β1、Gli2和IGF-II的表达水平以及细胞增殖、迁移和侵袭能力均升高。TGF-β1 在条件培养基中上调，并被发现参与促进 Huh-7 细胞的迁移、侵袭和 EMT。TGF-β1 信号的激活增加了 Gli2 的表达。Gli2 的敲低降低了 IGF-II 的表达，也逆转了条件培养基对 Huh-7 细胞迁移、侵袭和 EMT 的促进作用。TGF-β1/Gli2/IGF-II 信号通过激活 ERK1/2 信号通路促进 Huh-7 细胞的恶性表型。此外，TGF-β1 敲低减弱了 TAMs 对小鼠模型肿瘤生长的影响。

结论

TAMs分泌的TGF-β1通过Gli2/IGF-II/ERK1/2通路促进人肝癌Huh-7细胞的迁移、侵袭和EMT。