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Astragaloside IV suppresses migration and invasion of TGF-β1-induced human hepatoma HuH-7 cells by regulating Nrf2/HO-1 and TGF-β1/Smad3 pathways

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Abstract

Astragaloside IV (AS-IV), one of the major compounds extract from Astragalus membranaceus, has shown attractive anti-cancer effects in certain malignancies. Oxidative stress (OS) is considered as a crucial factor in promoting the progression of hepatocellular carcinoma (HCC). In response to OS, nuclear factor erythroid 2-related factor 2 (Nrf2) upregulates and induces heme oxygenase 1 (HO-1) to combat oxidative damages. The phosphorylation of the COOH-terminal of Smad3 (pSmad3C) activates p21 to resist HCC progression, while the phosphorylation of the linker region of Smad3 (pSmad3L) up-regulates c-Myc transcription to exert promoting effect towards HCC. This study aimed to explore whether AS-IV suppresses migration and invasion of human hepatoma HuH-7 cells by regulating Nrf2/HO-1 and TGF-β1/Smad3 pathways. HuH-7 cells were induced with TGF-β1 (9 or 40 pM) to establish HCC model in vitro and pretreated with AS-IV at different concentration (5, 10, and 20 μM) for 24 h. Cell proliferation, migration, invasion, and intracellular reactive oxygen species (ROS) of HuH-7 cells were measured. The expression of Nrf2, pSmad3C, Nrf2/pNrf2, HO-1, pSmad3C/3L, c-Myc, and p21 were detected. Exposure of HuH-7 cells to TGF-β1 enhanced the cell proliferation, migration, invasion, and ROS production. Pretreatment with AS-IV (5, 10, and 20 μM) significantly reduced the cell proliferation, migration, invasion, and ROS production in HuH-7 cells. Furthermore, AS-IV increased the expressions of Nrf2/pNrf2, HO-1, pSmad3C, and p21, meanwhile reduced the expressions of pSmad3L and c-Myc. In conclusion, our study suggested that AS-IV inhibit HuH-7 cells migration and invasion, which related to activate Nrf2/HO-1 pathway, up-regulation pSmad3C/p21 pathway, and down-regulation pSmad3L/c-Myc pathway. The present research supports the notion that AS-IV may be a latent agent for the treatment of HCC.

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Acknowledgements

We thank Dr. K.Matsuzaki for presenting us with rabbit polyclonal anti-pSmad3L (Ser208/213) antibody.

Funding

This work was financially supported by the National Natural Science Foundation of China (No 81874354; 82074073) and the Fund support by Anhui Medical University (No:2019GQFY13; 2020xkj035).

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Authors and Affiliations

  1. Department of Pharmacology, Key Laboratory of Anti-Inflammatory and Immunopharmacology, Ministry of Education, Anhui Medical University, Hefei, 230032, China

    Lili Li, Qin Wang, Yinghao He, Liangjie Sun, Yan Yang & Xiaonan Pang

  2. Department of Oncology, Second Affiliated Hospital of Anhui Medical University, Hefei, 230601, China

    Xiaonan Pang

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  1. Lili Li
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  2. Qin Wang
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  3. Yinghao He
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Contributions

LLL and YY conceived and designed the research. LLL conducted experiments and wrote the manuscript. QW contributed to new reagents and analytical tools. YHH provided the methodology and analyzed data. YY and XNP were responsible for the supervision and funding acquisition. XNP contributed to writing, reviewing, and editing the manuscript. All authors read and approved the manuscript, and all data were generated in-house and that no paper mill was used.

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Correspondence to Yan Yang or Xiaonan Pang.

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Li, L., Wang, Q., He, Y. et al. Astragaloside IV suppresses migration and invasion of TGF-β1-induced human hepatoma HuH-7 cells by regulating Nrf2/HO-1 and TGF-β1/Smad3 pathways. Naunyn-Schmiedeberg's Arch Pharmacol 395, 397–405 (2022). https://doi.org/10.1007/s00210-021-02199-8

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