Abstract
Java Ginger or Curcuma zanthorrhiza Roxb. has long gained focus among tribal people of Java, for its medicinal properties mainly against gynaecological challenges. The present study aims to identify the most potent phytocompound present in the extract and determine primary mode of action accountable for cytotoxic activity of Curcuma zanthorrhiza rhizome extract against HPV16-positive SiHa cervical cancer cells. The phytochemically-rich extract of rhizome (CZM) was capable to inhibit proliferation of target cells in a dose-dependent manner with an IC50 of 150 μg/ml. Dysregulation of intercellular antioxidant defence system resulted to surges in ROS and RNS level, increased calcium concentration and compromised mitochondrial membrane potential. Nucleus got affected, cell cycle dynamics got impaired while clonogenicity and migration ability diminished. Expression of viral oncogenes E7 and E6 decreased significantly. Accumulation of toxic cell metabolite and decrease in level of essential ones continued. Finally, alteration in PI3K/AKT/mTOR signalling route was followed by onset of autophagic cell death concomitant with the upregulated expression of Beclin1, Atg5-12 and LC3II. Curcumin and a novel crystal as well as few phyto-fractions were isolated by column chromatography. Of these, curcumin was found to be most potent in inducing cytotoxicity in SiHa while two other fractions also showed significant activity. Thus, CZM acted against SiHa cells by inducing autophagy that commences in compliance to the changes in PI3K/AKT/mTOR pathway mainly in response to oxidative stress. To the best of our knowledge this is the first report of Curcuma zanthorrhiza Roxb. inducing autophagy.
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Acknowledgements
SN would like to acknowledge Prof. Mamiyil Sabu, University of Calicut for kindly providing the rhizome of Curcuma zanthorrhiza Roxb. SAIF-IIT Bombay is acknowledged for HRLCMS analyses, University of Calcutta’s CRNN for allowing use of flow cytometer, DBT-IPLS facility for confocal microscope, UGC-UPE facility for gel-documentation unit. SN would like to thank all the former and present research scholars from Advanced Cell Biology Laboratory, Department of Botany, University of Calcutta, for their help. This work was supported by University Grants Commission-Universities with Potential for Excellence (UGC-UPE)-II, Grant number: UGC/144/UPE/ST1 (20.03.17). SN thanks the University Grants Commission, Government of India for her fellowship.
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SN and RK conceived the work. SN carried out the experiments and curated data. DP conducted NMR and helped with NMR and XRD analyses. AN conducted GCMS procedure and SN analysed the data. SN wrote the manuscript. RK assessed and confirmed the manuscript.
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Nath, S., Patra, D., Nag, A. et al. Sesquiterpenoid-rich Java Ginger rhizome extract prompts autophagic cell death in cervical cancer cell SiHa mainly by modulating cellular redox homeostasis. 3 Biotech 13, 8 (2023). https://doi.org/10.1007/s13205-022-03415-9
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DOI: https://doi.org/10.1007/s13205-022-03415-9