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Isorhamnetin exerts anti-tumor activity in DEN + CCl4-induced HCC mice

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Abstract

Background

Hepatocellular carcinoma (HCC) is the most prevalent type of liver cancer and the main cause of cancer death globally. The use of medicinal herbs as chemotherapeutic agents in cancer treatment is receiving attention as they possess no or minimum side effects. Isorhamnetin (IRN), a flavonoid, has been under attention for its anti-inflammatory and anti-proliferative properties in a number of cancers, including colorectal, skin, and lung cancers. However, the in vivo mechanism of isorhamnetin to suppress liver cancer has yet to be explored.

Methods and Result

HCC was induced by N-diethylnitrosamine (DEN) and carbon tetrachloride (CCL4) in Swiss albino mice. Isorhamnetin (100 mg/kg body weight) was given to examine its anti-tumor properties in HCC mice model. Histological analysis and liver function assays were performed to assess changes in liver anatomy. Probable molecular pathways were explored using immunoblot, qPCR, ELISA, and immunohistochemistry techniques. Isorhamnetin inhibited various pro-inflammatory cytokines to suppress cancer-inducing inflammation. Additionally, it regulated Akt and MAPKs to suppress Nrf2 signaling. Isorhamnetin activated PPAR-γ and autophagy while suppressing cell cycle progression in DEN + CCl4-administered mice. Additionally, isorhamnetin regulated various signaling pathways to suppress cell proliferation, metabolism, and epithelial–mesenchymal transition in HCC.

Conclusion

Regulating diverse cellular signaling pathways makes isorhamnetin a better anti-cancer chemotherapeutic candidate in HCC. Importantly, the anti-TNF-α properties of isorhamnetin could prove it a valuable therapeutic agent in sorafenib-resistant HCC patients. Additionally, anti-TGF-β properties of isorhamnetin could be utilized to reduce the EMT-inducing side effects of doxorubicin.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

ATG7:

Autophagy-related 7

Bad:

BCL2-associated agonist of cell death

CCl4 :

Carbon tetrachloride

C/EBP-δ:

CCAAT/enhancer-binding protein delta

DEN:

N-Diethylnitrosamine

Doxo:

Doxorubicin

ERK:

Extracellular signal-regulated kinase

EMT:

Epithelial–mesenchymal transition

GSK-3β:

Glycogen synthase kinase-3 beta:

HCC:

Hepatocellular carcinoma

HO-1:

Heme Oxygenase-1

HRP:

Horseradish peroxidise

IRN:

Isorhamnetin

JNK:

C-Jun N-terminal kinase

Keap1:

Kelch-like ECH-associated protein 1

Lamp2A:

Lysosomal associated membrane protein-2

MMP-9:

Matrix metalloproteinases 9

Mcl-1:

Myeloid cell leukemia-1

mTOR:

Mammalian target of rapamycin

Nrf2:

Nuclear factor erythroid 2-related factor 2

NSCLC:

Non-small cell lung carcinoma

PPAR-γ:

Peroxisome proliferator-activated receptor-gamma

ROS:

Reactive oxygen species

RSK:

Ribosomal S6 kinase

RUNX2:

Runt-related transcription factor 2

SOD:

Superoxide dismutase

SDS:

Sodium dodecyl sulfate

SVR:

Sustained virological response

STAT3:

Signal transducer and activator of transcription 3

TGF-β:

Transforming growth factor beta

TNF-α:

Tumor necrosis factor alpha

YAP1:

Yes-associated protein 1

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Acknowledgements

The authors are deeply grateful to Ms. Noyel Ghosh and Ms. Ankita Mandal for their valuable inputs during the preparation of the manuscript.

Funding

The authors declare that no funds, grants, or other support were received during the preparation of this manuscript.

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Authors and Affiliations

  1. Department of Life Sciences & Biotechnology, Jadavpur University, 188, Raja SC Mullick Road, Kolkata, 700032, India

    Sayanta Sarkar & Ratan Gachhui

  2. Division of Molecular Medicine, Bose Institute, P-1/12, CIT Scheme VII M, Kolkata, West Bengal, 700054, India

    Abhishek Kumar Das & Parames C. Sil

  3. Indian Association for the Cultivation of Science, 2A & 2B, Raja Subodh Chandra Mallick Rd, Jadavpur, Kolkata, West Bengal, 700032, India

    Semantee Bhattacharya

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Contributions

SS contributed to conceptualization, methodology, software, validation, data curation, formal analysis, investigation, writing of the original draft, visualization, and writing, reviewing, & editing of the manuscript. AKD contributed to methodology, validation, and writing, reviewing, & editing of the manuscript. SB contributed to conceptualization. RG contributed to conceptualization, validation, data curation, formal analysis, visualization, and writing, reviewing, & editing of the manuscript. PCS contributed to conceptualization, validation, data curation, formal analysis, visualization, and writing, reviewing, & editing of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Parames C. Sil.

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The authors have no relevant financial or non-financial interests to disclose.

Ethical approval

All the animal experiments were prosecuted according to the institutional ethical committee and with the permission of IAEC, CPCSEA (Committee for the Purpose of Control & Supervision on Experiments on Animals) and the Ministry of Environment and Forests, New Delhi, India [1796/GO/EReBiBt/S/14/CPCSEA].

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Sarkar, S., Das, A.K., Bhattacharya, S. et al. Isorhamnetin exerts anti-tumor activity in DEN + CCl4-induced HCC mice. Med Oncol 40, 188 (2023). https://doi.org/10.1007/s12032-023-02050-5

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