Skip to main content

Advertisement

SpringerLink
Log in

Berberine reverses epithelial-mesenchymal transition and modulates histone methylation in osteosarcoma cells

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Osteosarcoma is the most frequently occurring cancer in children as well as young adolescents and the metastatic forms worsen this condition to a further great extent. The metastatic dissemination of cancer cells is often acquired through a process of epithelial-mesenchymal transition (EMT). Since, phytochemicals have attracted intense interest in recent years due to their diverse pharmacological effects, in the present study, we investigated if berberine, a naturally occurring isoquinoline quaternary alkaloid, could modulate the EMT in osteosarcoma cells. Our experimental studies showed that berberine reduced cell viability, colony formation, wound healing ability and migration of osteosarcoma cells. Also, berberine significantly reduced the expression of matrix metalloproteinase (MMP)-2, suggesting its inhibitory action on the matrix metalloproteinases that are required for cancer cell invasion. The significant reduction in the expression of vimentin, N-cadherin, fibronectin and increased expression of E-cadherin further suggested its role in the inhibition of EMT in osteosarcoma cells. The downregulation of H3K27me3, as well as the decreased expression of the histone methyl transferase enzyme EZH2, further substantiated the fact that the plant alkaloid can be used as an epigenetic modulator in the treatment of osteosarcoma. In conclusion, our findings suggest that berberine inhibits proliferation and migration of osteosarcoma cells and most importantly reverses EMT along with modulation of key epigenetic regulators.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Mirabello L, Troisi RJ, Savage SA (2009) International osteosarcoma incidence patterns in children and adolescents, middle ages and elderly persons. Int J Cancer 125(1):229–234

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Mirabello L, Troisi RJ, Savage SA (2009) Osteosarcoma incidence and survival rates from 1973 to 2004: data from the Surveillance, Epidemiology, and End Results Program. Cancer: Interdiscip Int J Am Cancer Soc 115(7):1531–1543

    Article  Google Scholar 

  3. Kaste SC, Pratt CB, Cain AM, Jones-Wallace DJ, Rao BN (1999) Metastases detected at the time of diagnosis of primary pediatric extremity osteosarcoma at diagnosis: imaging features. Cancer 86(8):1602–1608

    Article  CAS  PubMed  Google Scholar 

  4. Huang X, Zhao J, Bai J, Shen H, Zhang B, Deng L, Sun C, Liu Y, Zhang J, Zheng J (2019) Risk and clinicopathological features of osteosarcoma metastasis to the lung: a population-based study. J Bone Oncol 16:100230

    Article  PubMed  PubMed Central  Google Scholar 

  5. Hunter KW, Crawford NP, Alsarraj J (2008) Mechanisms of metastasis. Breast Cancer Res 10, Article number: S2

  6. Gonzalez DM, Medici D (2014) Signaling mechanisms of the epithelial-mesenchymal transition. Sci Signal 7(344):re8

    Article  PubMed  PubMed Central  Google Scholar 

  7. Moreno-Bueno G, Portillo F, Cano A (2008) Transcriptional regulation of cell polarity in EMT and cancer. Oncogene 27(55):6958–6969

    Article  CAS  PubMed  Google Scholar 

  8. Serrano-Gomez SJ, Maziveyi M, Alahari SK (2016) Regulation of epithelial-mesenchymal transition through epigenetic and post-translational modifications. Mol Cancer 15(1), Article number: 18

  9. Hu Y, Dai M, Zheng Y, Wu J, Yu B, Zhang H, Kong W, Wu H, Yu X (2018) Epigenetic suppression of E-cadherin expression by Snail2 during the metastasis of colorectal cancer. Clin Epigenet 10(1), Article number: 154

  10. Zha L, Cao Q, Cui X, Li F, Liang H, Xue B, Shi H (2016) Epigenetic regulation of E-cadherin expression by the histone demethylase UTX in colon cancer cells. Med Oncol 33(3):21

    Article  PubMed  Google Scholar 

  11. Lv YF, Yan GN, Meng G, Zhang X, Guo QN (2015) Enhancer of zeste homolog 2 silencing inhibits tumor growth and lung metastasis in osteosarcoma. Sci Rep 5, Article number: 12999

  12. Chou AJ, Geller DS, Gorlick R (2008) Therapy for osteosarcoma. Pediatric Drugs 10(5):315–327

    Article  PubMed  Google Scholar 

  13. Neergheen VS, Bahorun T, Taylor EW, Jen LS, Aruoma OI (2010) Targeting specific cell signaling transduction pathways by dietary and medicinal phytochemicals in cancer chemoprevention. Toxicology 278(2):229–241

    Article  CAS  PubMed  Google Scholar 

  14. Hosseini A, Ghorbani A (2015) Cancer therapy with phytochemicals: evidence from clinical studies. Avicenna J Phytomed 5(2):84–97

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Vuddanda PR, Chakraborty S, Singh S (2010) Berberine: a potential phytochemical with multispectrum therapeutic activities. Expert Opin Investig Drugs 19(10):1297–1307

    Article  CAS  PubMed  Google Scholar 

  16. Cai C, Huang W, Zhou J, Wu Q, Huang Y, Zhao W, Zhang F, Wang Q, Zhang Y, Fang J (2019) An insight into the molecular mechanism of berberine towards multiple cancer types through systems pharmacology. Front Pharmacol 10:857

    Article  PubMed  PubMed Central  Google Scholar 

  17. Sun Y, Xun K, Wang Y, Chen X (2009) A systematic review of the anticancer properties of berberine, a natural product from Chinese herbs. Anticancer Drugs 20(9):757–769

    Article  CAS  PubMed  Google Scholar 

  18. Jin H, Jin X, Cao B, Wang W (2017) Berberine affects osteosarcoma via downregulating the caspase-1/IL-1β signaling axis. Oncol Rep 37(2):729–736

    Article  CAS  PubMed  Google Scholar 

  19. Liu Z, Liu Q, Xu B, Wu J, Guo C, Zhu F, Yang Q, Gao G, Gong Y, Shao C (2009) Berberine induces p53-dependent cell cycle arrest and apoptosis of human osteosarcoma cells by inflicting DNA damage. Mutat Res 662(1–2):75–83

    Article  CAS  PubMed  Google Scholar 

  20. Zhu Y, Ma N, Li HX, Tian L, Ba YF, Hao B (2014) Berberine induces apoptosis and DNA damage in MG–63 human osteosarcoma cells. Mol Med Rep 10(4):1734–1738

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Varshney R, Mishra R, Das N, Sircar D, Roy P (2019) A comparative analysis of various flavonoids in the regulation of obesity and diabetes: an in vitro and in vivo study. J Funct Foods 59:194–205

    Article  CAS  Google Scholar 

  22. Varshney R, Gupta S, Roy P (2017) Cytoprotective effect of kaempferol against palmitic acid-induced pancreatic β-cell death through modulation of autophagy via AMPK/mTOR signaling pathway. Mol Cell Endocrinol 448:1–20

    Article  CAS  PubMed  Google Scholar 

  23. Varshney R, Varshney R, Mishra R, Gupta S, Sircar D, Roy P (2018) Kaempferol alleviates palmitic acid-induced lipid stores, endoplasmic reticulum stress and pancreatic β-cell dysfunction through AMPK/mTOR-mediated lipophagy. J Nutr Biochem 57:212–227

    Article  CAS  PubMed  Google Scholar 

  24. Loh CY, Chai JY, Tang TF, Wong WF, Sethi G, Shanmugam MK, Chong PP, Looi CY (2019) The E-cadherin and N-cadherin switch in epithelial-to-mesenchymal transition: signaling, therapeutic implications, and challenges. Cells 8(10):1118

    Article  CAS  PubMed Central  Google Scholar 

  25. Tsai JH, Yang J (2013) Epithelial–mesenchymal plasticity in carcinoma metastasis. Genes Dev 27(20):2192–2206

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Yang G, Yuan J, Li K (2013) EMT transcription factors: implication in osteosarcoma. Med Oncol 30(4):697

    Article  PubMed  Google Scholar 

  27. Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100(1):57–70

    Article  CAS  PubMed  Google Scholar 

  28. Machesky LM (2008) Lamellipodia and filopodia in metastasis and invasion. FEBS Lett 582(14):2102–2111

    Article  CAS  PubMed  Google Scholar 

  29. Izdebska M, Zielińska W, Grzanka D, Gagat M (2018) The role of actin dynamics and actin-binding proteins expression in epithelial-to-mesenchymal transition and its association with cancer progression and evaluation of possible therapeutic targets. BioMed Res Int 2018, Article ID 4578373

  30. Chu SC, Yu CC, Hsu LS, Chen KS, Su MY, Chen PN (2014) Berberine reverses epithelial-to-mesenchymal transition and inhibits metastasis and tumor-induced angiogenesis in human cervical cancer cells. Mol Pharmacol 86(6):609–623

    Article  PubMed  Google Scholar 

  31. PosthumaDeBoer J, Witlox MA, Kaspers GJ, van Royen BJ (2011) Molecular alterations as target for therapy in metastatic osteosarcoma: a review of literature. Clin Exp Metas 28(5):493–503

    Article  CAS  Google Scholar 

  32. Kleiner DE, Stetler-Stevenson WG (1999) Matrix metalloproteinases and metastasis. Cancer Chemother Pharmacol 43(1):S42–S51

    Article  CAS  PubMed  Google Scholar 

  33. Bjørnland K, Flatmark K, Pettersen S, Aaasen AO, Fodstad O, Melandsmo GM (2005) Matrix metalloproteinases participate in osteosarcoma invasion. J Surg Res 127(2):151–156

    Article  PubMed  Google Scholar 

  34. Dave B, Mittal V, Tan NM, Chang JC (2012) Epithelial-mesenchymal transition, cancer stem cells and treatment resistance. Breast Cancer Res 14(1), Article number: 202

  35. Yang H, Zhang Y, Zhou Z, Jiang X, Shen A (2014) Transcription factor Snai1–1 induces osteosarcoma invasion and metastasis by inhibiting E–cadherin expression. Oncol Lett 8(1):193–197

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Kim K, Lu Z, Hay ED (2002) Direct evidence for a role of β-catenin/LEF-1 signaling pathway in induction of EMT. Cell Biol Int 26(5):463–476

    Article  CAS  PubMed  Google Scholar 

  37. Chen C, Zhao M, Tian A, Zhang X, Yao Z, Ma X (2015) Aberrant activation of Wnt/β-catenin signaling drives proliferation of bone sarcoma cells. Oncotarget 6(19):17570–17583

    Article  PubMed  PubMed Central  Google Scholar 

  38. Lin CH, Ji T, Chen CF, Hoang BH (2014) Wnt signaling in osteosarcoma. Adv Exp Med Biol 804:33–45

    Article  CAS  PubMed  Google Scholar 

  39. Gottardi CJ, Wong E, Gumbiner BM (2001) E-cadherin suppresses cellular transformation by inhibiting β-catenin signaling in an adhesion-independent manner. J Cell Biol 153(5):1049–1060

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Feng H, Tillman H, Wu G, Davidoff AM, Yang J (2018) Frequent epigenetic alterations in polycomb repressive complex 2 in osteosarcoma cell lines. Oncotarget 9(43):27087–27091

    Article  PubMed  PubMed Central  Google Scholar 

  41. Liu D, Meng X, Wu D, Qiu Z, Luo H (2019) A natural isoquinoline alkaloid with antitumor activity: studies of the biological activities of berberine. Front Pharmacol 10:9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Wang Z, Liu Y, Xue Y, Hu H, Ye J, Li X, Lu Z, Meng F, Liang S (2016) Berberine acts as a putative epigenetic modulator by affecting the histone code. Toxicol In Vitro 36:10–17

    Article  CAS  PubMed  Google Scholar 

  43. Chen S, Chen X, Li Y, Zhang F, Yang S, Mo KL, Mo X, Ding Y (2015) Inhibitory effect of berberine on zeste homolog 2 (Ezh2) enhancement in human esophageal cell lines. Trop J Pharm Res 14(9):1589–1595

    CAS  Google Scholar 

Download references

Acknowledgements

The authors are highly grateful to Michigan Diagnostics, Royal Oak, MI, USA for the kind gift of immune-detection reagent for western blot analysis.

Funding

This study was funded by research grants from Ministry of Human Resource and Development, Government of India and Indian Institute of Technology Roorkee, Roorkee, as research fellowship to RM.

Author information

Authors and Affiliations

  1. Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247 667, India

    Rutusmita Mishra, Sandip Nathani, Ritu Varshney & Partha Roy

  2. Plant Molecular Biology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247 667, India

    Debabrata Sircar

Authors
  1. Rutusmita Mishra
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sandip Nathani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ritu Varshney
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Debabrata Sircar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Partha Roy
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

RM: Conceptualization, Data curation. Investigation, Formal analysis, Validation, Writing—original draft. SN: Data curation, Formal analysis, Writing—original draft. RV: Data curation, Formal analysis, Writing—original draft. DS: Resources, and Visualization. PR: Project administration, Resources, Supervision, Conceptualization, Visualization, Validation, Writing—Review & Editing.

Corresponding author

Correspondence to Partha Roy.

Ethics declarations

Conflicts of interest

RM declares that she has no conflict of interest. SN declares that he has no conflict of interest. RV declares that she has no conflict of interest. DS declares that he has no conflict of interest. PR declares that he has no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 24 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mishra, R., Nathani, S., Varshney, R. et al. Berberine reverses epithelial-mesenchymal transition and modulates histone methylation in osteosarcoma cells. Mol Biol Rep 47, 8499–8511 (2020). https://doi.org/10.1007/s11033-020-05892-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-020-05892-8

Keywords

Search

Navigation