Humanized disulfide-stabilized diabody against fibroblast growth factor-2 inhibits PD-L1 expression and epithelial-mesenchymal transition in hepatoma cells through STAT3
Humanized disulfide-stabilized diabody against fibroblast growth factor-2 inhibits PD-L1 expression and epithelial-mesenchymal transition in hepatoma cells through STAT3
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DOI：
10.1002/iub.2766
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发表时间：
2023-07-25
期刊：
影响因子：
4.6
通讯作者：
Deng，Ning
中科院分区：
文献类型：
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作者：
Sun，Huamin;Song，Xinran;Deng，Ning
成纤维细胞生长因子2 (FGF2) 在肿瘤血管生成中发挥重要作用。人源化二硫键稳定的抗成纤维细胞生长因子-2双链抗体（抗FGF2 ds-Diabody）是一种具有良好组织渗透性和低免疫原性的小分子抗体，在肿瘤靶向治疗中具有潜力。本研究旨在探讨抗 FGF2 ds-Diabody 对肝细胞癌 (HCC) 细胞中程序性死亡配体 1 (PD-L1) 迁移和表达的影响。抗 FGF2 ds-Diabody 在甲醇诱导下表达，并用 Ni2+-亲和层析纯化。 CCK-8 测定和集落形成测定证实，抗 FGF2 ds-Diabody 显着抑制 SK-Hep1 和 HepG2 细胞的细胞活力和增殖。 Western blot 检测表明，抗 FGF2 ds-Diabody 通过抑制 AKT 和 MAPK 的磷酸化激活来抑制 SK-Hep1 和 HepG2 细胞的增殖。 Transwell和Western blot检测结果表明，抗FGF2 ds-Diabody通过影响上皮间质转化（EMT）过程来抑制SK-Hep1和HepG2细胞的迁移和侵袭。同时，抗FGF2 ds-Diabody抑制PD-L1的表达，STAT3参与了这一过程。 RT-PCR 和 Western blot 分析表明，成纤维细胞生长因子受体 4 抑制剂 1 (FGFR4-IN-1) 抑制 PD-L1 的表达，而 STAT3 过表达则逆转了这种抑制作用。此外，STAT3的过表达促进了迁移和侵袭，并恢复了抗FGF2 ds-Diabody对EMT的抑制作用。总之，抗FGF2 ds-Diabody可以通过FGF2/FGFR4/STAT3轴抑制肝癌细胞PD-L1和EMT的表达。这些结果表明抗FGF2 ds-Diabody在抑制肝细胞癌的转移和免疫逃逸方面具有潜在的临床应用。
Fibroblast growth factor 2 (FGF2) plays an important role in tumor angiogenesis. Humanized disulfide‐stable double‐chain antibody against fibroblast growth factor‐2 (anti‐FGF2 ds‐Diabody) is a small molecule antibody with good tissue permeability and low immunogenicity, which has potential in tumor‐targeted therapy. This study intended to investigate the effect of anti‐FGF2 ds‐Diabody on the migration and expression of programmed death‐ligand1 (PD‐L1) in hepatocellular carcinoma (HCC) cells. The anti‐FGF2 ds‐Diabody was expressed under methanol induction and purified with Ni2+‐affinity chromatography. Anti‐FGF2 ds‐Diabody significantly inhibited cell viability and proliferation in SK‐Hep1 and HepG2 cells as confirmed by CCK‐8 assays and colony formation assays. Western blot assays indicated that the proliferation of SK‐Hep1 and HepG2 cells was inhibited by anti‐FGF2 ds‐Diabody through inhibiting the phosphorylation activation of AKT and MAPK. The results of transwell and western blot assays showed that the migration and invasion of SK‐Hep1 and HepG2 cells were suppressed by anti‐FGF2 ds‐Diabody by affecting the epithelial‐mesenchymal transition (EMT) process. Meanwhile, anti‐FGF2 ds‐Diabody inhibited the expression of PD‐L1, and STAT3 participated in this process. Analysis of RT‐PCR and Western blot suggested that fibroblast growth factor receptor 4 inhibitor 1 (FGFR4‐IN‐1) suppressed the expression of PD‐L1, while STAT3 overexpression reversed this inhibitory effect. In addition, overexpression of STAT3 promoted migration and invasion and restored the suppressive effect of anti‐FGF2 ds‐Diabody on EMT. In conclusion, anti‐FGF2 ds‐Diabody could inhibit the expression of PD‐L1 and EMT of hepatoma cells through FGF2/FGFR4/STAT3 axis. These results suggested that anti‐FGF2 ds‐Diabody has potential clinical application in inhibiting metastasis and immune escape of hepatocellular carcinoma.