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Micromorphological observation of HLE cells under knockdown of Fascin using LV-SEM

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Abstract

Liver cancer is one of the most prevalent cancers in Japan with hepatocellular carcinoma (HCC) as the major histological subtype. Successful novel treatments for HCC have been reported; however, recurrences or metastasis may occur, which results in poor prognoses and high mortality of HCC patients. Fascin, an actin-bundling protein, regulates cell adhesion, migration, and invasion. Its overexpression positively correlates with poor prognosis of malignant tumors, and Fascin is considered as one of the tumor biomarkers and therapeutic target proteins. In this study, we attempted to reveal the relationship between Fascin and HCC using HLE, one of the human HCC cell lines. We performed the study with classical immunocytochemistry and recently developed techniques, such as wound-healing assay, spheroid cultivation, and low-vacuum scanning electron microscopy (LV-SEM). Non-Fascin-knockdown (FKD) cell spheroid had a regular spherical appearance with tight cell–cell connections, while FKD cell spheroid had an irregular shape with loose cell–cell connections. Cells of non-FKD spheroid presented fibrous protrusions on the cell surface, contrarily, cells of FKD spheroids showed bulbous-shaped protrusions. Morphological observation of FKD and non-FKD HLE spheroids were performed using LV-SEM. Our study may help to reveal the roles of Fascin in the process of HCC formation and its malignancy.

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Authors agree that all materials described in the present manuscript, including all relevant raw data, are freely available to any researchers who may wish to reuse or reanalyze them.

References

  1. National Cancer Center Japan. https://www.ncc.go.jp/jp/index.html

  2. Pu J, Zhang Y, Wang A, Qin Z, Zhuo C, Li W, Xu Z, Tang Q, Wang J, Wei H (2021) ADORA2A-AS1 restricts hepatocellular carcinoma progression via binding HuR and repressing FSCN1/AKT axis. Front Oncol 11:754835

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Llovet JM, Montal R, Sia D, Finn RS (2018) Molecular therapies and precision medicine for hepatocellular carcinoma. Nat Rev Clin Oncol 15:599–616

    Article  PubMed  Google Scholar 

  4. Xing R, Gao J, Cui Q, Wang Q (2021) Strategies to improve the antitumor effect of immunotherapy for hepatocellular carcinoma. Front Immunol 12:783236

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Han S, Lim JY, Cho K, Lee HW, Park JY, Ro SW, Kim KS, Seo HR, Kim DY (2022) Anti-cancer effects of YAP inhibitor (CA3) in combination with sorafenib against hepatocellular carcinoma (HCC) in patient-derived multicellular tumor spheroid models (MCTS). Cancers 14:2733

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Yamamoto Y, Hayashi Y, Sakaki H, Murakami I (2021) Fascin-1 is associated with recurrence in solitary fibrous tumor/hemangiopericytoma. Mol Clin Oncol 15:199

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Hayashi Y, Osanai M, Lee G-H (2011) Fascin-1 expression correlates with repression of E-cadherin expression in hepatocellular carcinoma cells and augments their invasiveness in combination with matrix metalloproteinases. Cancer Sci 102:1228–1235

    Article  CAS  PubMed  Google Scholar 

  8. Wang CQ, Tang CH, Chang HT, Li XN, Zhao YM, Su CM, Hu GN, Tao Zhang T, Sun XX, Zeng Y, Du Z, Wang Y, Huang GF (2016) Fascin-1 as a novel diagnostic marker of triple-negative breast cancer. Med 5:1983–1988

    CAS  Google Scholar 

  9. Arlt MJ, Kuzmanov A, Snedeker JG, Fuchs B, Silvan U, Sabile AA (2019) Fascin-1 enhances experimental osteosarcoma tumor formation and metastasis and is related to poor patient outcome. BMC Cancer 19:83

    Article  PubMed  PubMed Central  Google Scholar 

  10. Abbasi A, Noroozinia F, Anvar S, Hosseinzadeh S, Mokhtari S (2019) Fascin overexpression is associated with higher grades of breast cancer. Pol J Pathol 70:264–268

    Article  PubMed  Google Scholar 

  11. Yamamoto Y, Hayashi Y, Sakaki H, Murakami I (2022) Evaluation of clinical and immunohistochemical factors relating to melanoma metastasis: potential roles of nestin and fascin in melanoma. Diagnostics 12:219

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Lin S, Taylor M, Singh P, Yang S (2021) How does fascin promote cancer metastasis? FEBS J 288:1434–1446

    Article  CAS  PubMed  Google Scholar 

  13. Liu H, Zhang Y, Li L, Cao J, Guo Y, Wu Y, Gao W (2021) Fascin actin-bundling protein 1 in human cancer: promising biomarker or therapeutic target? Mol Ther Oncolytics 20:240–264

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Yamamoto Y, Hayashi Y, Sakaki H, Murakami I (2023) Downregulation of fascin induces collective cell migration in triple-negative breast cancer. Oncol Rep. https://doi.org/10.3892/or.2023.8587

    Article  PubMed  PubMed Central  Google Scholar 

  15. Hayashi Y, Toda K, Saibara T, Okamoto S, Osanai M, Enzan H, Lee GH (2008) Expression of fascin-1, an actin-bundling protein, in migrating hepatoblasts during rat liver development. Cell Tissue Res 334:219–226

    Article  CAS  PubMed  Google Scholar 

  16. Hayashi Y, Yamamoto Y, Murakami I (2022) Fascin expression persists with fibronectin in embryonic rat hepatoblasts. Med Mol Morphol 55:100–109

    Article  CAS  PubMed  Google Scholar 

  17. Lu W, Kang Y (2019) Epithelial-mesenchymal plasticity in cancer progression and metastasis. Dev Cell 49:361–374

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Ma Y, Zhang P, Wang F, Yang J, Yang Z, Qin H (2010) The relationship between early embryo development and tumourigenesis. J Cell Mol Med 14:2697–2701

    Article  PubMed  PubMed Central  Google Scholar 

  19. Huang Y, Hong W, Wei X (2022) The molecular mechanisms and therapeutic strategies of EMT in tumor progression and metastasis. J Hematol Oncol 15:129

    Article  PubMed  PubMed Central  Google Scholar 

  20. Di Masi A, Viganotti M, Antoccia A, Magrelli A, Salvatore M, Azzalin G, Tosto F, Lorenzetti S, Maranghi F, Mantovani A, Macino G, Tanzarella C, Taruscio D (2010) Characterization of HUH6, HEP3B, HEPG2 and HLE liver cancer cell lines by WNT/Q- catenin pathway, microrna expression and protein expression profile. Cell Mol Biol 56:1299–1317

    Google Scholar 

  21. Iguchi T, Aishima S, Umeda K, Sanefuji K, Fujita N, Sugimachi K, Gion T, Taketomi A, Maehara Y, Tsuneyoshi M (2009) Fascin expression in progression and prognosis of hepatocellular carcinoma. J Surg Oncol 100:575–579

    Article  PubMed  Google Scholar 

  22. Huang X, Ji J, Xue H, Zhang F, Han X, Cai Y, Zhang J, Ji G (2012) Fascin and cortactin expression is correlated with a poor prognosis in hepatocellular carcinoma. Eur J Gastroenterol Hepatol 24:633–639

    Article  CAS  PubMed  Google Scholar 

  23. Zhang Y, Lu Y, Zhang C, Huang D, Wu W, Zhang Y, Shen J, Cai Y, Chen W, Yao W (2018) FSCN-1 increases doxorubicin resistance in hepatocellular carcinoma through promotion of epithelial-mesenchymal transition. Int J Oncol 52:1455–1464

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Martinotti S, Ranzato E (2020) Scratch wound healing assay. Methods Mol biol 2109:225–229

    Article  CAS  PubMed  Google Scholar 

  25. Grada A, Marta OV, Francisco PC, Zaidal O, Vincent F (2017) Research techniques made simple: analysis of collective cell migration using the wound healing assay. J Invest Dermatol 137:e11–e16

    Article  CAS  PubMed  Google Scholar 

  26. Alfonso García SL, Mira Uribe LM, Castaño López S, Parada-Sanchez MT, Arboleda-Toro D (2022) Ultrastructural characterization of human gingival fibroblasts in 3D culture. Cells 11:3647

    Article  PubMed  PubMed Central  Google Scholar 

  27. Huang Z, Yu P, Tang J (2020) Characterization of triple-negative breast cancer MDA-MB-231 cell spheroid model. Onco Targets Ther 13:5395–5405

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Marshall J (2011) Transwell® invasion assays. Methods Mol Biol 769:97–110

    Article  CAS  PubMed  Google Scholar 

  29. Lamb MC, Tootle TL (2020) Fascin in cell migration: more than an actin bundling protein. Biology 9:403

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We would like to thank Cherish Warden for English editing. We also appreciate the LVSEM Study Group of Renal Biopsy and Hitachi High-Tech for a provision of the LVSEM as a part of the grant research No. 006 of the LVSEM Study Group of Renal Biopsy.

Funding

No funding was received for the present study.

Author information

Authors and Affiliations

  1. Department of Pathology, School of Medicine, Kochi University, 185-1, Kohasu, Oko-Cho, Nankoku, 783-8505, Japan

    Yoshihiro Hayashi & Ichiro Murakami

  2. Equipment of Support Planning Office, Kochi University, 185-1, Kohasu, Oko-Cho, Nankoku, 783-8505, Japan

    Yoshihiro Hayashi

  3. Department of Diagnostic Pathology, Kochi University Hospital, Kochi University, 185-1, Kohasu, Oko-Cho, Nankoku, 783-8505, Japan

    Yumiko Yamamoto & Ichiro Murakami

Authors
  1. Yoshihiro Hayashi
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  2. Yumiko Yamamoto
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Contributions

YH and YY conceived, planned, and carried out the experiments. YH is the main contributor for performing the experiment and YY is for writing the manuscript. IM supervised the creation of the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Yumiko Yamamoto.

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Hayashi, Y., Yamamoto, Y. & Murakami, I. Micromorphological observation of HLE cells under knockdown of Fascin using LV-SEM. Med Mol Morphol 56, 257–265 (2023). https://doi.org/10.1007/s00795-023-00362-z

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  • DOI: https://doi.org/10.1007/s00795-023-00362-z

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