Homeopathy 2022; 111(04): 288-300
DOI: 10.1055/s-0042-1743565
Original Research Article

Computational and In Vitro Approaches to Elucidate the Anti-cancer Effects of Arnica montana in Hormone-Dependent Breast Cancer

Nilanjana Basu
1   Amity Institute of Molecular Medicine and Stem Cell Research, Amity University, Noida, Uttar Pradesh, India
Priyanka Narad
2   Amity Institute of Biotechnology, Amity University, Noida, Uttar Pradesh, India
Manni Luthra Guptasarma
3   Department of Immunopathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India
Chanderdeep Tandon
4   Amity University Mohali, Punjab, India
Bhudev Chandra Das
1   Amity Institute of Molecular Medicine and Stem Cell Research, Amity University, Noida, Uttar Pradesh, India
1   Amity Institute of Molecular Medicine and Stem Cell Research, Amity University, Noida, Uttar Pradesh, India
4   Amity University Mohali, Punjab, India
› Author Affiliations


Background Breast cancer is the most common cancer in women worldwide. Use of homeopathic medicines for the treatment of cancers has increased in the last several years. Arnica montana is an anti-inflammatory homeopathic medicine used in traumatic conditions and because of this property we performed investigations for its potential as a chemotherapeutic agent against breast cancer.

Methods An ethanolic extract of Arnica montana (mother tincture, MT), prepared according to the Homoeopathic Pharmacopoeia of India, was characterized by gas chromatography–mass spectroscopy (GC–MS), followed by computational (in silico) analysis using molecular docking, to identify specific compounds that can bind and modulate the activity of key proteins involved in breast cancer survival and progression. To validate the in silico findings, in a controlled experiment breast cancer cells (MCF7) were treated in vitro with Arnica montana and the cytotoxic effects assessed by flowcytometry, fluorescence microscopy, scratch assay, clonogenic potential and gene expression analysis.

Results Phytochemical characterization of ethanolic extract of Arn MT by GC–MS allowed identification of several compounds. Caryophyllene oxide and 7-hydroxycadalene were selected for molecular docking studies, based on their potential drug-like properties. These compounds displayed selective binding affinity to some of the recognized target proteins of breast cancer, which included estrogen receptor alpha (ERα), progesterone receptor (PR), epidermal growth factor receptor (EGFR), mTOR (mechanistic target of rapamycin) and E-cadherin. In vitro studies revealed induction of apoptosis in MCF7 cells following treatment with Arn MT. Furthermore, treatment with Arn MT revealed its ability to inhibit migration and colony forming abilities of the cancer cells.

Conclusion Considering the apoptotic and anti-migratory effects of Arnica montana in breast cancer cells in vitro, there is a need for this medicine to be further validated in an in vivo model.

Authors' Contributions

N.B. conceptualized the study, performed the investigation and the formal analysis, and was a major contributor in writing the manuscript. P.N. performed the investigation and formal analysis, and edited the manuscript. M.L.G., C.T., and B.C.D. performed formal analysis. S.T. conceptualized, designed and supervised the study, and edited the manuscript.

Supplementary Material

Publication History

Received: 02 September 2021

Accepted: 12 November 2021

Article published online:
05 July 2022

© 2022. Faculty of Homeopathy. This article is published by Thieme.

Georg Thieme Verlag KG
Rüdigerstraße 14, 70469 Stuttgart, Germany

  • References

  • 1 World Health Organization. Breast cancer now most common form of cancer: WHO taking action. Available at: https://www.who.int/news/item/03-02-2021-breast-cancer-now-most-common-form-of-cancer-who-taking-action
  • 2 Torre LA, Sauer AMG, Chen Jr MS, Kagawa-Singer M, Jemal A, Siegel RL. Cancer statistics for Asian Americans, Native Hawaiians, and Pacific Islanders, 2016: converging incidence in males and females. CA Cancer J Clin 2016; 66: 182-202
  • 3 Stark GB, Grandel S, Spilker G. Tissue suction of the male and female breast. Aesthetic Plast Surg 1992; 16: 317-324
  • 4 Blamey R, Collins J, Crosignani PG. et al. Hormones and Breast Cancer. Vol. 10, Human Reproduction Update. Oxford University Press; 2004: 281-293
  • 5 Tiash S, Chowdhury E. Growth factor receptors: promising drug targets in cancer. J Cancer Metastasis Treat 2015; 1: 190-200
  • 6 Alzahrani AS. PI3K/Akt/mTOR inhibitors in cancer: At the bench and bedside. Semin Cancer Biol 2019; 59: 125-132
  • 7 Wu Y, Zhang Z, Cenciarini ME. et al. Tamoxifen resistance in breast cancer is regulated by the EZH2–ERa–GREB1 transcriptional axis. Cancer Res 2018; 78: 671-684
  • 8 Arun A, Ansari MI, Popli P. et al. New piperidine derivative DTPEP acts as dual-acting anti-breast cancer agent by targeting ERα and downregulating PI3K/Akt-PKCα leading to caspase-dependent apoptosis. Cell Prolif 2018; 51: e12501
  • 9 Migliaccio A, Piccolo D, Castoria G. et al. Activation of the Src/p21ras/Erk pathway by progesterone receptor via cross-talk with estrogen receptor. EMBO J 1998; 17: 2008-2018
  • 10 Caner A, Asik E, Ozpolat B. SRC signaling in cancer and tumor microenvironment. Adv Exp Med Biol 2021; 1270: 57-71
  • 11 Fuentes N, Silveyra P. Estrogen receptor signaling mechanisms. Adv Protein Chem Struct Biol 2019; 116: 135-170
  • 12 Conneely OM, Jericevic BM, Lydon JP. Progesterone receptors in mammary gland development and tumorigenesis. J Mammary Gland Biol Neoplasia 2003; 8: 205-214
  • 13 Sharma D, Kumar S, Narasimhan B. Estrogen alpha receptor antagonists for the treatment of breast cancer: a review. Chem Cent J 2018; 12: 107
  • 14 Lei JT, Anurag M, Haricharan S, Gou X, Ellis MJ. Endocrine therapy resistance: new insights. Breast 2019; 48: S26-S30
  • 15 Jeong Y, Bae SY, You D. et al. EGFR is a therapeutic target in hormone receptor-positive breast cancer. Cell Physiol Biochem 2019; 53: 805-819
  • 16 Bachelot T, Bourgier C, Cropet C. et al. Randomized phase II trial of Everolimus in combination with tamoxifen in patients with hormone receptor-positive, human epidermal growth factor receptor 2-negative metastatic breast cancer with prior exposure to aromatase inhibitors: a GINECO study. J Clin Oncol 2012; 30: 2718-2724
  • 17 Royce M, Bachelot T, Villanueva C. et al. Everolimus plus endocrine therapy for postmenopausal women with estrogen receptor-positive, human epidermal growth factor receptor 2-negative advanced breast cancer: a clinical trial. JAMA Oncol 2018; 4: 977-984
  • 18 Pećina-Šlaus N. Tumor suppressor gene E-cadherin and its role in normal and malignant cells. Cancer Cell Int 2003; 3: 17
  • 19 Bracke ME, Charlier C, Bruyneel EA, Labit C, Mareel MM, Castronovo V. Tamoxifen restores the E-cadherin function in human breast cancer MCF-7/6 cells and suppresses their invasive phenotype. Cancer Res 1994; 54: 4607-4609
  • 20 Cukaci C, Freissmuth M, Mann C, Marti J, Sperl V. Against all odds–the persistent popularity of homeopathy. Wien Klin Wochenschr 2020; 132: 232-242
  • 21 Relton C, Cooper K, Viksveen P, Fibert P, Thomas K. Prevalence of homeopathy use by the general population worldwide: A systematic review. Homeopathy 2017; 106: 69-78
  • 22 Bhattacharya TS, Maitra P, Bera D. et al. Investigation of the origin of voltage generation in potentized homeopathic medicine through Raman spectroscopy. Homeopathy 2019; 108: 121-127
  • 23 Banerjee A, Pathak S, Biswas SJ. et al. Chelidonium majus 30C and 200C in induced hepato-toxicity in rats. Homeopathy 2010; 99: 167-176
  • 24 Bellavite P, Magnani P, Zanolin E, Conforti A. Homeopathic doses of Gelsemium sempervirens improve the behavior of mice in response to novel environments. Evid Based Complement Alternat Med 2011; 2011: 362517
  • 25 Saha S, Bhattacharjee P, Mukherjee S. et al. Contribution of the ROS–p53 feedback loop in thuja-induced apoptosis of mammary epithelial carcinoma cells. Oncol Rep 2014; 31: 1589-1598
  • 26 Chikramane PS, Suresh AK, Bellare JR, Kane SG. Extreme homeopathic dilutions retain starting materials: a nanoparticulate perspective. Homeopathy 2010; 99: 231-242
  • 27 Kriplani P, Guarve K, Baghael US. Arnica montana L.—a plant of healing: review. J Pharm Pharmacol 2017; 69: 925-945
  • 28 Macêdo SB, Ferreira LR, Perazzo FF, Carvalho JC. Anti-inflammatory activity of Arnica montana 6cH: preclinical study in animals. Homeopathy 2004; 93: 84-87
  • 29 Kawakami AP, Sato C, Cardoso TN, Bonamin LV. Inflammatory process modulation by homeopathic Arnica montana 6CH: the role of individual variation. Evid Based Complement Alternat Med 2011; 2011: 917541
  • 30 Marzotto M, Arruda-Silva F, Bellavite P. Fibronectin gene up-regulation by Arnica montana in human macrophages: validation by real-time polymerase chain reaction assay. Homeopathy 2020; 109: 140-145
  • 31 Olioso D, Marzotto M, Bonafini C, Brizzi M, Bellavite P. Arnica montana effects on gene expression in a human macrophage cell line. Evaluation by quantitative real-time PCR. Homeopathy 2016; 105: 131-147
  • 32 Hahnemann S. Organon of Medicine. 5th and 6th ed.. B. Jain Publishers Pvt. Ltd.; 2008: 67
  • 33 Hughes JP, Rees S, Kalindjian SB, Philpott KL. Principles of early drug discovery. Br J Pharmacol 2011; 162: 1239-1249
  • 34 Boericke W. Pocket Manual of Homeopathic Materia Medica. Reprinted. Kandern, Germany: Narayana Publishers; 2013: 92-94
  • 35 Rigby JE, Morris JA, Lavelle J, Stewart M, Gatrell AC. Can physical trauma cause breast cancer?. Eur J Cancer Prev 2002; 11: 307-311
  • 36 Cotesta M, Buonomo OC, De Majo A. et al. Breast trauma and triple-negative hemorrhagic cystic carcinoma: management and treatment. Am J Case Rep 2020; 21: e925014
  • 37 Thomas P, Sullivan D-S. Breast mass detected after blunt chest trauma. Accessed June 30, 2021: https://www.ajronline.org/doi/pdfplus/10.2214/ajr.171.1.9648761
  • 38 Fisusi FA, Akala EO. Drug combinations in breast cancer therapy. Pharm Nanotechnol 2019; 7: 3-23
  • 39 Park KR, Nam D, Yun HM. et al. β-Caryophyllene oxide inhibits growth and induces apoptosis through the suppression of PI3K/AKT/mTOR/S6K1 pathways and ROS-mediated MAPKs activation. Cancer Lett 2011; 312: 178-188
  • 40 Egas V, Millán E, Collado JA. et al. Effect of natural and semi-synthetic Cadinanes from Heterotheca inuloides on NF-κB, Nrf2 and STAT3 signaling pathways and evaluation of their in vitro cytotoxicity in human cancer cell lines. Bioorg Med Chem 2017; 25: 3135-3147
  • 41 Boonsri S, Karalai C, Ponglimanont C, Chantrapromma S, Kanjana-Opas A. Cytotoxic and antibacterial sesquiterpenes from Thespesia populnea . J Nat Prod 2008; 71: 1173-1177
  • 42 Palma G, Frasci G, Chirico A. et al. Triple negative breast cancer: looking for the missing link between biology and treatments. Oncotarget 2015; 6: 26560-26574
  • 43 Saxton RA, Sabatini DM. mTOR signaling in growth, metabolism, and disease. Cell; 2017. 168. 960-976
  • 44 Cosulich SC, Worrall V, Hedge PJ, Green S, Clarke PR. Regulation of apoptosis by BH3 domains in a cell-free system. Curr Biol 1997; 7: 913-920
  • 45 Giotakis AI, Kontos CK, Manolopoulos LD, Sismanis A, Konstadoulakis MM, Scorilas A. High BAX/BCL2 mRNA ratio predicts favorable prognosis in laryngeal squamous cell carcinoma, particularly in patients with negative lymph nodes at the time of diagnosis. Clin Biochem 2016; 49: 890-896
  • 46 Köhler T, Schill C, Deininger MW. et al. High Bad and Bax mRNA expression correlate with negative outcome in acute myeloid leukemia (AML). Leukemia 2002; 16: 22-29
  • 47 Pijuan J, Barceló C, Moreno DF. et al. In vitro cell migration, invasion, and adhesion assays: from cell imaging to data analysis. Front Cell Dev Biol 2019; 7: 107
  • 48 Rafehi H, Orlowski C, Georgiadis GT, Ververis K, El-Osta A, Karagiannis TC. Clonogenic assay: adherent cells. J Vis Exp 2011; 49
  • 49 Adhikary A, Chakraborty S, Mazumdar M. et al. Inhibition of epithelial to mesenchymal transition by E-cadherin up-regulation via repression of slug transcription and inhibition of E-cadherin degradation: dual role of scaffold/matrix attachment region-binding protein 1 (SMAR1) in breast cancer cells. J Biol Chem 2014; 289: 25431-25444
  • 50 Zhang X, Zhang B, Liu J. et al. Mechanisms of Gefitinib-mediated reversal of tamoxifen resistance in MCF-7 breast cancer cells by inducing ERα re-expression. Sci Rep 2015; 5: 1-7
  • 51 Osborne CK, Neven P, Dirix LY. et al. Gefitinib or placebo in combination with tamoxifen in patients with hormone receptor-positive metastatic breast cancer: a randomized phase II study. Clin Cancer Res 2011; 17: 1147-1159
  • 52 Wang Q, Gun M, Hong XY. Induced tamoxifen resistance is mediated by increased methylation of E-cadherin in estrogen receptor-expressing breast cancer cells. Sci Rep 2019; 9: 14140