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Cited by 18Medicinal Plant ResearchPhytochemical Studies and BioactivitiesPhytochemicals and Medicinal Plants

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Tiliacora racemosa leaves induce oxidative stress mediated DNA damage leading to G2/M phase arrest and apoptosis in cervical cancer cells SiHa

Pal, Asmita, Sengupta, Soumee, Kundu, Rita·2021·Journal of Ethnopharmacology
DOI 10.1016/j.jep.2020.113686
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Abstract

Ethnopharmacological relevance The Menispermaceae plant Tiliacora racemosa is immensely popular in Indian traditional Ayurvedic medicine as "Krishnavetra" for its remarkable anti-cancerous property, and is commonly used by tribal population for the treatment of skin infections, snake bites and filariasis. Aim of the study This present study intends to identify the modus operandi behind the cytotoxic activity of Tiliacora racemosa leaves in cervical cancer cells SiHa. Focus has been instilled in the ability of the plant extract to target multiple signaling pathways leading to cell cycle arrest and cell death in SiHa cells, followed by a pharmacological characterization to identify the bioactive principle. Materials and methods T. racemosa leaves extracted in methanol, ethyl acetate, hexane and aqueous solvent were screened for cytotoxicity in HeLa, SiHa, C33A (cervical cancer cells) and HEK cells by MTT assay. SiHa cells were treated with the most potent extract (TRM). Cellular morphology, clonogenic and wound healing potential, presence of intracellular ROS and NO, lipid peroxidation, activity of cellular antioxidants (SOD, CAT, GSH), DNA damage detection by comet assay and localisation of γ-H2AX foci, intracellular expression of PARP-1, Bax/Bcl2 and caspase-3, loss in mitochondrial membrane potential by JC1 (flow cytometry) and Rh123 (microscopy), cell cycle analysis, Annexin-FITC assay, AO/EtBr microscopy and apoptotic proteome profiling were undertaken in the treated cells. All the related proteins were studied by immunoblots. Effect of NAC (ROS-scavenger) on cell viability, DNA damage and apoptosis were studied. Phytochemical characterization of all TR extracts was followed by LC-MS analysis of TRM and isolated alkaloid of TR was assessed for cytotoxicity. Results The methanol extract of T. racemosa (TRM) rich in bisbenzylisoquinoline and other alkaloids impeded the proliferation of cervical cancer cells SiHa in vitro through disruption of cellular redox homeostasis caused by increase in cellular ROS and NO with concomitant decrease in the cellular antioxidants. Double-stranded DNA damage was noted from γH2AX foci accumulation and Parp-1 activation leading to ATM-Chk2-p53 pathway arresting the cells at G2/M-phase through cyclin B1 inhibition. The mitochondrial membrane potential was also disturbed leading to caspase-3 dependent apoptotic induction by both extrinsic and intrinsic pathway. Immunoblots show TRM also inhibited PI3K/Akt and NFκB pathway. NAC pre-treatment rescued the cell viability proving DNA damage and apoptosis to be direct consequences of ROS overproduction. Lastly, the therapeutic potential of T. racemosa is was hypothesized to be possibly derived from its alkaloid content. Conclusion This study proves the age old ethnnopharmacological anticancer role of T. racemosa. The leaf extracts inhibited the anomalous proliferation of SiHa cells by virtue of G2/M-phase cell cycle arrest and apoptotic cell death. Oxidative stress mediated double stranded DNA damage paved the way towards apoptotic cell death through multiple routes, including PI3K/Akt/NFκB pathway. The abundant alkaloid content of T. racemosa was denoted as the probable responsible cytotoxic principle.

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Metadata

Authors
Pal, A., Sengupta, S., & Kundu, R.
Affiliations
University of Calcutta (IN)
Corresponding author
Kundu, Rita
ORCID
Venue
Journal of Ethnopharmacology
Publisher
Elsevier BV
Published
2021-04
Volume
269
Pages
113686
ISSN
0378-8741 (ISSN-L)
DOI
10.1016/j.jep.2020.113686
OpenAlex
W3111335901
Cited by
18
References
3
Citations by year
2026: 2 · 2025: 4 · 2024: 4 · 2023: 3 · 2022: 3 · 2021: 2
FWCI
1.29
Type
journal article
Language
en
Update notices
erratum (2024-05-23)
Open access
closed
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • Journal of Ethnopharmacology (publishedVersion)
  • PubMed (publishedVersion)
Funders
University Grants Commission
Topics
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Medicinal Plant Research
  • Physical Sciences › Chemistry › Organic Chemistry › Phytochemical Studies and Bioactivities
  • Health Sciences › Medicine › Complementary and alternative medicine › Phytochemicals and Medicinal Plants
Keywords
cervical cancer, SiHa cells, oxidative stress, DNA damage, G2/M cell cycle arrest, apoptosis, bisbenzylisoquinoline alkaloids, reactive oxygen species, nitric oxide, redox homeostasis, γH2AX foci, p53, ATM-Chk2 pathway, Bcl2/Bax, caspase-3
MeSH terms
Antineoplastic Agents, Phytogenic, Cell Division, Cell Survival, Uterine Cervical Neoplasms, DNA Damage, Dose-Response Relationship, Drug, Female, HeLa Cells, Humans, G2 Phase, Oxidative Stress, Plant Leaves, Menispermaceae, HEK293 Cells

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CoChat. (2026). Citation check: Tiliacora racemosa leaves induce oxidative stress mediated DNA damage leading to G2/M phase arrest and apoptosis in cervical cancer cells SiHa. https://app.cochat.ai/papers/10.1016/j.jep.2020.113686
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Pal, A., Sengupta, S., & Kundu, R. (2021). Tiliacora racemosa leaves induce oxidative stress mediated DNA damage leading to G2/M phase arrest and apoptosis in cervical cancer cells SiHa. Journal of Ethnopharmacology. https://doi.org/10.1016/j.jep.2020.113686