CoChat
Sign Up
Cited by 18Medicinal Plant ResearchPhytochemical Studies and BioactivitiesPhytochemicals and Medicinal Plants

The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.

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
Open this paper in CoChat — free account

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.

Overview

Integrity signals

Registry

Crossref · 10.1016/j.jep.2020.113686

Cites

3 of 3 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Verification

Claims the paper makes

We couldn’t retrieve this paper’s full text, so its claims and self-check weren’t run. The registry and citation checks still apply.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Citations

Cites

Research works printed but not linked: 52

  1. Ajanal, 2012, Estimation of total alkaloid in Chitrakadivati by UV-Spectrophotometer, Ancient Sci. Life, 31, 198, 10.4103/0257-7941.107361 · DOI
  2. Anjaneyulu, 1969, Alkaloids of Tiliacora racemosa Colebr, Tetrahedron, 25, 3091, 10.1016/S0040-4020(01)82842-8 · DOI
  3. Bai, 2009, Akt-mediated regulation of NFkappaB and the essentialness of NFkappaB for the oncogenicity of PI3K and Akt, Int. J. Canc., 125, 2863, 10.1002/ijc.24748 · DOI · Paper page
  4. Bray, 2018, Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries, CA Canc. J. Clin., 68, 394, 10.3322/caac.21492 · DOI · Paper page
  5. Broadhurst, 1978, Analysis of condensed tannins using acidified vanillin, J. Sci. Food Agric., 29, 788, 10.1002/jsfa.2740290908 · DOI
  6. Brüne, 2003, Nitric oxide: NO apoptosis or turning it ON?, Cell Death Differ., 10, 864, 10.1038/sj.cdd.4401261 · DOI
  7. Carneiro, 2020, Targeting apoptosis in cancer therapy, Nat. Rev. Clin. Oncol., 10.1038/s41571-020-0341-y · DOI
  8. Chakraborty, 2004, Cytotoxic effect of root extract of Tiliacora racemosa and oil of Semecarpus anacardium nut in human tumour cells, Phytother Res., 18, 595, 10.1002/ptr.1501 · DOI
  9. Chandra, 2007, Cytosolic accumulation of HSP60 during apoptosis with or without apparent mitochondrial release: evidence that its pro-apoptotic or pro-survival functions involve differential interactions with caspase-3, J. Biol. Chem., 282, 31289, 10.1074/jbc.M702777200 · DOI
  10. Chandrakanthan, 2014, Antibacterial, antioxidant and antiproliferative activities of solvent extracts of Tiliacora acuminata, Int. J. Pharm. Pharmaceut. Sci., 6, 398
  11. Christ, 1960, Zur serienmaessigen Bestimmung des Gehaltes an Flavonol-Derivaten in Drogen, Arch. Pharmazie, 293, 1033, 10.1002/ardp.19602931202 · DOI
  12. Chung, 2017, Sinularin induces DNA damage, G2/M phase arrest, and apoptosis in human hepatocellular carcinoma cells, BMC Compl. Alternative Med., 17, 62, 10.1186/s12906-017-1583-9 · DOI
  13. Cunin, 2016, Clusterin facilitates apoptotic cell clearance and prevents apoptotic cell induced auto-immune responses, Cell Death Dis., 7, e2215, 10.1038/cddis.2016.113 · DOI
  14. Du, 2000, Smac, a mitochondrial protein that promotes cytochrome c–dependent caspase activation by eliminating IAP inhibition, Cell, 102, 33, 10.1016/S0092-8674(00)00008-8 · DOI · Paper page
  15. Esposti, 2002, The roles of Bid, Apoptosis, 7, 433, 10.1023/A:1020035124855 · DOI
  16. Ghorai, 2012, Estimation of total terpenoids concentration in plant tissues using a monoterpene, linalool as standard reagent, Protoc. Exch., 10.1038/protex.2012.055 · DOI
  17. Ghosh, 2014, Ethnobotanical use of plants as living fence in and around radhanagar village of jhargram block, paschim medinipur district, West Bengal, Indian J. Appl. Pure Biol., 29, 223
  18. Gillis, 2009, p21Cip1/WAF1 mediates cyclin B1 degradation in response to DNA damage, Cell Cycle, 8, 253, 10.4161/cc.8.2.7550 · DOI
  19. Gottlieb, 2002, Cross-talk between Akt, p53 and Mdm2: possible implications for the regulation of apoptosis, Oncogene, 21, 1299, 10.1038/sj.onc.1205181 · DOI
  20. Hirao, 2000, DNA damage-induced activation of p53 by the checkpoint kinase Chk2, Science, 287, 1824, 10.1126/science.287.5459.1824 · DOI
  21. Huang, 2019, Functional characterization of HSP60-survivin interaction between mitochondria and cytosol in cancer cells, Cell, 9, 23, 10.3390/cells9010023 · DOI
  22. Hurley, 2002, DNA and its associated processes as targets for cancer therapy, Nature, 2, 188
  23. Janeklang, 2014, In vitro and in vivo antitumor activity of tiliacorinine in human cholangiocarcinoma, Asian Pac. J. Cancer Prev. APJCP, 15, 7473, 10.7314/APJCP.2014.15.17.7473 · DOI
  24. Johnson, 2004, Survivin: a bifunctional inhibitor of apoptosis protein, Vet. Pathol., 41, 599, 10.1354/vp.41-6-599 · DOI
  25. Kasof, 2001, Livin, a novel inhibitor of apoptosis protein family member, J. Biol. Chem., 276, 3238, 10.1074/jbc.M003670200 · DOI
  26. Kilic, 2007, Role of hypoxia inducible factor-1 alpha in modulation of apoptosis resistance, Oncogene, 26, 2027, 10.1038/sj.onc.1210008 · DOI
  27. Kumar, 2017, Evaluation of antibacterial, antioxidant and nootropic activities of Tiliacora racemosa Colebr. leaves: in vitro and in vivo approach, Biomed. Pharmacother., 86, 662, 10.1016/j.biopha.2016.12.030 · DOI
  28. Liu, 1997, DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis, Cell, 89, 175, 10.1016/S0092-8674(00)80197-X · DOI
  29. Mathur, 2020, Cancer statistics, 2020: report from national cancer registry programme, India, JCO Global Oncol., 6, 1063, 10.1200/GO.20.00122 · DOI · Paper page
  30. McCord, 1969, Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein), J. Biol. Chem., 244, 6049, 10.1016/S0021-9258(18)63504-5 · DOI · Paper page
  31. Olive, 1990, Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the "comet" assay, Radiat. Res., 122, 86, 10.2307/3577587 · DOI
  32. Pan, 2019, Cinobufagin induces cell cycle arrest at the G2/M phase and promotes apoptosis in malignant melanoma cells, Front Oncol, 9, 853, 10.3389/fonc.2019.00853 · DOI
  33. Payne, 2008, p27kip1 deficiency impairs G2/M arrest in response to DNA damage, leading to an increase in genetic instability, Mol. Cell Biol., 28, 258, 10.1128/MCB.01536-07 · DOI
  34. Pelicano, 2004, ROS stress in cancer cells and therapeutic implications, Drug Resist. Updates, 7, 97, 10.1016/j.drup.2004.01.004 · DOI
  35. Pugazhenthi, 2000, Akt/protein kinase B up-regulates Bcl-2 expression through cAMP-response element-binding protein, J. Biol. Chem., 275, 10.1074/jbc.275.15.10761 · DOI · Paper page
  36. Rashid, 2013, Taurine ameliorate alloxan induced oxidative stress and intrinsic apoptotic pathway in the hepatic tissue of diabetic rats, Food Chem. Toxicol., 51, 317, 10.1016/j.fct.2012.10.007 · DOI
  37. Ruprecht, 2019, Improved identification of DNA double strand breaks: γ-H2AX-epitope visualization by confocal microscopy and 3D reconstructed images, Radiat. Environ. Biophys., 58, 295, 10.1007/s00411-019-00778-1 · DOI
  38. Roy, 1997, The c‐IAP‐1 and c‐IAP‐2 proteins are direct inhibitors of specific caspases, EMBO J., 16, 6914, 10.1093/emboj/16.23.6914 · DOI
  39. Seal, 2002, (+)-Tiliarine, a selective in vitro inhibitor of human melanoma cells, Phytother Res., 16, 596, 10.1002/ptr.993 · DOI
  40. Scott, 2005, XIAP inhibits caspase-3 and -7 using two binding sites: evolutionarily conserved mechanism of IAPs, EMBO J., 24, 645, 10.1038/sj.emboj.7600544 · DOI
  41. Shangguan, 2014, Induction of G2/M phase cell cycle arrest and apoptosis by ginsenoside Rf in human osteosarcoma MG-63 cells through the mitochondrial pathway, Oncol. Rep., 31, 305, 10.3892/or.2013.2815 · DOI
  42. Shiozaki, 2003, Mechanism of XIAP-mediated inhibition of caspase-9, Mol. Cell., 11, 519, 10.1016/S1097-2765(03)00054-6 · DOI
  43. Siede, 1996, Cloning and characterization of RAD17, a gene controlling cell cycle responses to DNA damage in Saccharomyces cerevisiae, Nucleic Acids Res., 24, 1669, 10.1093/nar/24.9.1669 · DOI
  44. Singleton, 1965, Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents, Am. J. Enol. Vitic., 16, 144, 10.5344/ajev.1965.16.3.144 · DOI · Paper page
  45. Snyder, 2009, Nitric oxide induces cell death by regulating anti-apoptotic BCL-2 family members, PloS One, 4, 10.1371/journal.pone.0007059 · DOI
  46. Stehlik, 1998, Cytokine induced expression of porcine inhibitor of apoptosis protein (iap) family member is regulated by NF-κB, Biochem. Biophys. Res. Commun., 243, 827, 10.1006/bbrc.1998.8185 · DOI
  47. Takahashi, 2005, Does γH2AX foci formation depend on the presence of DNA double strand breaks?, Canc. Lett., 229, 171, 10.1016/j.canlet.2005.07.016 · DOI · Paper page
  48. Toshiyuki, 1995, Tumor suppressor p53 is a direct transcriptional activator of the human bax gene, Cell, 80, 293, 10.1016/0092-8674(95)90412-3 · DOI · Paper page
  49. Tsujimoto, 1998, Role of Bcl-2 family proteins in apoptosis: apoptosomes or mitochondria?, Gene Cell., 3, 697, 10.1046/j.1365-2443.1998.00223.x · DOI
  50. Wang, 2014, HSP27, 70 and 90, anti-apoptotic proteins, in clinical cancer therapy (review), Int. J. Oncol., 45, 18, 10.3892/ijo.2014.2399 · DOI
  51. Wang, 2019, Drug resistance and combating drug resistance in cancer, Canc. Drug Resist, 2, 141
  52. Zou, 1999, An APAF-1.cytochrome c multimeric complex is a functional apoptosome that activates procaspase-9, J. Biol. Chem., 274, 10.1074/jbc.274.17.11549 · DOI

Cited by

Sorted by citations

See all 18

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

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review