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Volume 25, Issue 3
  • ISSN: 1566-5240
  • E-ISSN: 1875-5666

Abstract

Background

The emergence of treatment resistance has hindered the efficacy of targeted therapies used to treat patients with hepatocellular carcinoma (HCC).

Objective

This study aimed to explore the mechanism of organoids constructed from lenvatinib-resistant HCC cells.

Methods

Hep3B cell and human HCC organoids were cultured and identified using hematoxylin and eosin staining and Immunohistochemistry. Lenvatinib-sensitive/ resistant Hep3B cells were constructed using lenvatinib (0, 0.1, 1, and 10 μM) and lenvatinib (0, 1, 10, and 100 μM). qRT-PCR and flow cytometry were utilized to deter-mine HCC stem cell markers CD44, CD90, and CD133 expressions. Transcriptome sequencing was performed on organoids. Western blot evaluated Notch pathway-related proteins (NOTCH1 and Jagged) expressions. Furthermore, DAPT, an inhibitor of the Notch pathway, was used to investigate the effects of lenvatinib on resistance or stemness in organoids and human HCC tissues.

Results

The organoids were successfully cultivated. With the increase of lenvatinib concentration, sensitive cell organoids were markedly degraded and ATP activity was gradually decreased, while there was no significant change in ATP activity of resistant cell organoids. CD44 expressions were elevated after lenvatinib treatment compared with the control group. KEGG showed that lenvatinib treatment of organoids constructed from Hep3B cells mainly activated the Notch pathway. Compared with the control group, NOTCH1 and Jagged expressions elevated, and ATP activity decreased after lenvatinib treatment. However, ATP activity was notably decreased after DAPT treatment. Moreover, DAPT inhibited lenvatinib resistance and the increase in the expressions of CD44 caused by lenvatinib. Besides, 100 μM lenvatinib significantly inhibited the growth and ATP activity of human HCC organoids, and DAPT increased the inhibitory effect of lenvatinib.

Conclusion

Lenvatinib regulated resistance and stemness in organoids the Notch pathway.

© 2025 Bentham Science Publishers
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2025-03-01
2025-06-24

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Study on the Mechanism of Notch Pathway Mediates the Role of Lenvatinib-resistant Hepatocellular Carcinoma Based on Organoids
Current Molecular Medicine 25, 343 (2025); https://doi.org/10.2174/0115665240268201231213095302
/content/journals/cmm/10.2174/0115665240268201231213095302
/content/journals/cmm/10.2174/0115665240268201231213095302
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  • Article Type:     Research Article
Keyword(s): drug resistance; hepatocellular carcinoma; Lenvatinib; notch pathway; organoid; stemness

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