Liensinine induces autophagy and apoptosis in hepatocellular carcinoma via reactive oxygen species-mediated inhibition of the PI3K/AKT/mTOR pathway

  • Tissue Cell. 2026 Jun 12:103:103696. doi: 10.1016/j.tice.2026.103696.
Xia Wang  1 ,  Jian Chang  2 ,  Xiaoli Zhang  3 ,  Yueli Hu  4 ,  Geqiong Xiao  5
Affiliations
  • 1. Department of Traditional Chinese Medicine, East Lake Street community health service center, Shaoxing 312000, China.
  • 2. Department of Medical Oncology, Affiliated Hospital of Shaoxing University (The Shaoxing Municipal Hospital), Shaoxing 312000, China.
  • 3. Department of hospital sense, Shaoxing People's Hospital, Shaoxing 312000, China.
  • 4. Department of Gynecology and obstetrics, Keqiao District Maternal and Child Health Hospital, Shaoxing 312000, China.
  • 5. Department of Medical Oncology, Affiliated Hospital of Shaoxing University (The Shaoxing Municipal Hospital), Shaoxing 312000, China. Electronic address: [email protected].
PMID: 42314534 DOI: 10.1016/j.tice.2026.103696
Abstract

Background: Liensinine (Lien), a bisbenzylisoquinoline alkaloid with anti-cancer properties, has shown potential in the treatment of hepatocellular carcinoma (HCC). This study aimed to elucidate the molecular mechanisms underlying the anti-HCC effects of Lien.

Methods: HepG2 cells were subcutaneously injected into nude mice, followed by intraperitoneal administration of Lien at 5 or 20 mg/kg. Tumor volume and weight were measured. The effects of Lien on Apoptosis and Autophagy in HCC were evaluated using Western blotting, immunofluorescence, transmission electron microscopy (TEM), and immunohistochemistry. In vitro, HepG2 cells were treated with 20 or 80 μM Lien. Alterations in Apoptosis, Autophagy, Reactive Oxygen Species (ROS) levels, and the PI3K/Akt/mTOR pathway were assessed using flow cytometry, Western blotting, and TEM. Furthermore, the mechanisms were investigated using the Autophagy inhibitor hydroxychloroquine (HCQ), ROS scavenger N-acetylcysteine (NAC), and PI3K/Akt pathway activator Recilisib.

Results: Lien significantly reduced tumor volume and weight and induced Apoptosis in HCC-bearing mice. Additionally, Lien notably enhanced Autophagy, as evidenced by increased autophagosome formation, elevated Beclin-1 and LC3-II/I levels, and decreased p62 expression. In vitro, Lien markedly inhibited HepG2 cell proliferation and promoted both Apoptosis and autophagic flux. These effects were partially attenuated by HCQ. Furthermore, NAC reversed Lien-induced changes in Apoptosis and Autophagy in HepG2 cells, accompanied by partial restoration of PI3K/Akt/mTOR pathway activity. Recilisib attenuated the inhibitory effects of Lien on the PI3K/Akt/mTOR pathway and partially reversed its effects on cell proliferation, Autophagy, and Apoptosis.

Conclusion: Lien exerts anti-HCC effects by promoting Autophagy and Apoptosis through ROS-mediated inhibition of the PI3K/Akt/mTOR pathway.

Keywords
Autophagy; Hepatocellular carcinoma; Liensinine; PI3K/AKT/mTOR pathway; Reactive oxygen species.
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