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Efficacy of punarnavine in restraining organ-specific tumour progression in 4T1-induced murine breast tumour model

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Abstract

Most of the breast cancer deaths occur when cancer cells depart from their tumour of origin and spread systemically and colonise distant organs. The present study was to find out whether punarnavine, the quinolizidine alkaloid, with already proven antimetastatic effect on spontaneous B16F10 pulmonary metastasis has got any effect on a drastic organ-specific breast cancer spread. For the study, we selected a syngenic mouse 4T1 breast tumour model that mimics stage four of human breast cancer. The metastatic progression of 4T1 to lymph nodes, lungs, and liver was reduced by punarnavine (40 mg/kg body weight) administration in BALB/c mice. This was evident from the histopathology of these organs as well as from the reduction in the metastatic cell density of cultured 6-thioguanine-resistant 4T1 cells in the punarnavine-treated group compared to the control group. There was also a significant (p < 0.0001) inhibition of the primary breast tumour growth in the orthotopic site of induction with a simultaneous increase (p < 0.0001) in the life span of treated animals. The assessment of biochemical parameters such as hydroxyproline, hexosamine, uronic acid, sialic acid and γ-glutamyl transferase and the analysis of various cytokines VEGF, IL-1β, TNF-α and GM-CSF showed a similar pattern of reduction in punarnavine (p < 0.0001) treated group compared to the control group. The gene expression study revealed the inhibitory effect of punarnavine on the major genes MMP-2, MMP-9, TIMP-1, TIMP-2 and VEGF involved in the metastatic process. These findings undeniably proved the potential of this quinolizidine alkaloid in combating breast tumour development and its progression in the studied murine model.

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Abbreviations

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Foetal bovine serum

HBSS:

Hanks balanced salt solution

ELISA:

Enzyme-linked immunosorbent assay

IL-1β:

Interleukin-1β

TNF-α:

Tumour necrosis factor-α

GMCSF:

Granulocyte monocyte colony-stimulating factor

VEGF:

Vascular endothelial growth factor

MMP:

Matrix metalloproteinase

TIMP:

Tissue inhibitor of matrix metalloproteinase

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

TNBC:

Triple-negative breast cancer

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Acknowledgements

The authors express gratitude to the Council of Scientific and Industrial Research (CSIR), Government of India for the senior research fellowship provided to Ms. Gilcy George K.

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Authors and Affiliations

  1. Department of Immunology, Amala Cancer Research Centre (Affiliated to the University of Calicut), Thrissur, Kerala, 680555, India

    Gilcy George Kallivalappil & Girija Kuttan

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  1. Gilcy George Kallivalappil
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  2. Girija Kuttan
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Correspondence to Girija Kuttan.

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George Kallivalappil, G., Kuttan, G. Efficacy of punarnavine in restraining organ-specific tumour progression in 4T1-induced murine breast tumour model. Inflammopharmacol 27, 701–712 (2019). https://doi.org/10.1007/s10787-018-0490-0

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  • DOI: https://doi.org/10.1007/s10787-018-0490-0

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