Tumor-associated macrophages promote human hepatoma Huh-7 cell migration and invasion through the Gli2/IGF-II/ERK1/2 axis by secreting TGF-β1
Tumor-associated macrophages promote human hepatoma Huh-7 cell migration and invasion through the Gli2/IGF-II/ERK1/2 axis by secreting TGF-β1
复制标题
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。
DOI：
10.1080/15384047.2020.1824478
复制
发表时间：
2020-10-22
影响因子：
3.6
通讯作者：
Shi, Chao
中科院分区：
文献类型：
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作者：
Liu, Mei;Zhong, Yuan-Bin;Shi, Chao
目的探讨TAMs通过Gli 2/IGF-II/ERK 1/2通路影响人肝癌细胞Huh-7恶性表型的能力。方法采用流式细胞术和ELISA法对TAMs进行鉴定。用TAMs条件培养液（TAMs-CM）处理Huh-7细胞，CCK-8法、Transwell法和划痕法检测TAMs对Huh-7细胞增殖、迁移和侵袭能力的影响。采用RT-qPCR和western blot检测ERK 1/2通路和上皮-间充质转化（EMT）过程中TGF-β 1、Gli 2、IGF-II及相关蛋白的水平。将TAM注射到Huh-7细胞中，以探讨TAM在肿瘤生长中的作用。结果经TAMs-CM处理后，Huh-7细胞TGF-β 1、Gli 2和IGF-II的表达水平升高，细胞增殖、迁移和侵袭能力增强。TGF-β 1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-β 1信号的激活增加了Gli 2的表达。Gli 2基因的敲除降低了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-β 1/Gli 2/IGF-II信号传导被证明通过激活ERK 1/2信号传导途径促进Huh-7细胞的恶性表型。此外，TGF-β 1敲低减弱了TAM对小鼠模型中肿瘤生长的影响。结论TAMs分泌的TGF-β 1通过Gli 2/IGF-II/ERK 1/2途径促进人肝癌Huh-7细胞的迁移、侵袭和EMT。
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-beta 1, 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-beta 1, Gli2 and IGF-II and the cell proliferation, migration and invasion abilities were elevated in Huh-7 cells treated with TAMs-CM. TGF-beta 1 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-beta 1 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-beta 1/Gli2/IGF-II signaling was shown to promote the malignant phenotype of Huh-7 cells by activating the ERK1/2 signaling pathway. Further, TGF-beta 1 knockdown attenuated the influence of TAMs on tumor growth in mouse model. Conclusion The TGF-beta 1 secreted by TAMs promotes the migration, invasion and EMT of human hepatoma Huh-7 cells through the Gli2/IGF-II/ERK1/2 pathway.