Skip to main content
SpringerLink
Log in

Naringenin attenuates cell viability and migration of C6 glioblastoma cell line: a possible role of hedgehog signaling pathway

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

Abstract

Objective

Gliomas are the most prevalent type of malignant primary brain tumors. Despite the availability of several treatment modalities, these tumors have poor prognostic features. Aberrant Hedgehog (Hh) signaling has been found to be implicated in the development of numerous malignancies including gliomas. Naringenin appears to have anti-proliferative and anti-cancer properties. However, there is no report describing its effects via the Hh signaling pathway on the C6 glioblastoma cell line. The current study was set to examine the anti-cancer effects of naringenin on C6 cells in order to determine the effect of this compound on the Hh signaling pathway.

Methods

The anti-proliferative and apoptotic effects of naringenin against C6 and 3T3 fibroblast cells were measured by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay and annexin-V/PI dual staining assay, respectively. The effect of naringenin on the migration of C6 cells was evaluated by the migration scratch assay. To assess the anti-cancer effect of naringenin on the Hh signaling pathway, the expression of Gli-1, Smo, and Sufu at protein levels in C6 cells was analyzed using western blotting.

Results

The obtained data indicated that naringenin exerted higher cytotoxicity against C6 cells (IC50 value of 114 ± 3.4 µg/mL) than normal 3T3 fibroblasts (IC50 value of 290 ± 7 µg/mL). Naringenin (114 µg/mL) also induced stronger apoptotic effects on C6 cells than 3T3 cells after 24 h of incubation. Furthermore, naringenin at a concentration of 114 µg/mL and a lower concentration of 60 µg/mL inhibited the migration of the C6 cell line. In addition, naringenin at a concentration of 114 µg/mL significantly decreased the expression of Gli-1 and Smo and elevated the expression of Sufu at the protein level in the C6 cell line.

Conclusion

These data represent that naringenin may have a potential effect on the management of the proliferation and metastasis of malignant gliomas by inhibiting the Hh signaling pathway.

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

Similar content being viewed by others

References

  1. Rynkeviciene R et al (2019) Non-coding RNAs in glioma. Cancers 11(1):17

    Article  CAS  Google Scholar 

  2. Melamed JR et al (2018) Investigating the role of Hedgehog/GLI1 signaling in glioblastoma cell response to temozolomide. Oncotarget 9(43):27000

    Article  Google Scholar 

  3. Petrova R, Joyner AL (2014) Roles for Hedgehog signaling in adult organ homeostasis and repair. Development 141(18):3445–3457

    Article  CAS  Google Scholar 

  4. Hanna A, Shevde LA (2016) Hedgehog signaling: modulation of cancer properies and tumor mircroenvironment. Mol Cancer 15(1):1–14

    Article  Google Scholar 

  5. Jeng KS et al (2019) Sonic Hedgehog signaling pathway as a potential target to inhibit the progression of hepatocellular carcinoma. Oncol Lett 18(5):4377–4384

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Du WZ et al (2013) Curcumin suppresses malignant glioma cells growth and induces apoptosis by inhibition of SHH/GLI 1 signaling pathway in vitro and vivo. CNS Neurosci Ther 19(12):926–936

    Article  CAS  Google Scholar 

  7. de Souza PO et al (2018) Anticancer activity of flavonoids isolated from Achyrocline satureioides in gliomas cell lines. Toxicol In Vitro 51:23–33

    Article  Google Scholar 

  8. Zanotto-Filho A et al (2013) Curcumin-loaded lipid-core nanocapsules as a strategy to improve pharmacological efficacy of curcumin in glioma treatment. Eur J Pharm Biopharm 83(2):156–167

    Article  CAS  Google Scholar 

  9. Bao C et al (2018) Regulation of hedgehog signaling in cancer by natural and dietary compounds. Mol Nutr Food Res 62(1):1700621

    Article  Google Scholar 

  10. Tian H et al (2007) A high throughput drug screen based on fluorescence resonance energy transfer (FRET) for anticancer activity of compounds from herbal medicine. Br J Pharmacol 150(3):321–334

    Article  CAS  Google Scholar 

  11. Chang HL et al (2017) Naringenin inhibits migration of lung cancer cells via the inhibition of matrix metalloproteinases-2 and-9. Exp Ther Med 13(2):739–744

    Article  CAS  Google Scholar 

  12. Salehi B et al (2019) The therapeutic potential of naringenin: a review of clinical trials. Pharmaceuticals 12(1):11

    Article  CAS  Google Scholar 

  13. Zhao Z et al (2019) Naringenin inhibits migration of breast cancer cells via inflammatory and apoptosis cell signaling pathways. Inflammopharmacology 27(5):1021–1036

    Article  CAS  Google Scholar 

  14. Aktas HG, Akgun T (2018) Naringenin inhibits prostate cancer metastasis by blocking voltage-gated sodium channels. Biomed Pharmacother 106:770–775

    Article  Google Scholar 

  15. Kumar R, Tiku AB (2020) Naringenin suppresses chemically induced skin cancer in two-stage skin carcinogenesis mouse model. Nutr Cancer 72(6):976–983

    Article  CAS  Google Scholar 

  16. Song HM et al (2015) Anti-proliferative effect of naringenin through p38-dependent downregulation of cyclin D1 in human colorectal cancer cells. Biomol Ther 23(4):339

    Article  CAS  Google Scholar 

  17. Chen YY et al (2019) Naringenin inhibited migration and invasion of glioblastoma cells through multiple mechanisms. Environ Toxicol 34(3):233–239

    Article  CAS  Google Scholar 

  18. Kanno S-I et al (2006) Naringenin-induced apoptosis via activation of NF-κB and necrosis involving the loss of ATP in human promyeloleukemia HL-60 cells. Toxicol Lett 166(2):131–139

    Article  CAS  Google Scholar 

  19. Wang B-D et al (2006) Synthesis, characterization, cytotoxic activities, and DNA-binding properties of the La (III) complex with Naringenin Schiff-base. Bioorg Med Chem 14(6):1880–1888

    Article  CAS  Google Scholar 

  20. Arul D, Subramanian P (2013) Naringenin (citrus flavonone) induces growth inhibition, cell cycle arrest and apoptosis in human hepatocellular carcinoma cells. Pathol Oncol Res 19(4):763–770

    Article  CAS  Google Scholar 

  21. Jin C-Y et al (2009) Naringenin-induced apoptosis is attenuated by Bcl-2 but restored by the small molecule Bcl-2 inhibitor, HA 14–1, in human leukemia U937 cells. Toxicol In Vitro 23(2):259–265

    Article  CAS  Google Scholar 

  22. Tundis R et al (2011) In vitro cytotoxic activity of extracts and isolated constituents of Salvia leriifolia Benth. against a panel of human cancer cell lines. Chem Biodivers 8(6):1152–1162

    Article  CAS  Google Scholar 

  23. Sabarinathan D, Mahalakshmi P, Vanisree AJ (2011) Naringenin, a flavanone inhibits the proliferation of cerebrally implanted C6 glioma cells in rats. Chem Biol Interact 189(1–2):26–36

    Article  CAS  Google Scholar 

  24. Stompor M, Uram Ł, Podgórski R (2017) In vitro effect of 8-prenylnaringenin and naringenin on fibroblasts and glioblastoma cells-cellular accumulation and cytotoxicity. Molecules 22(7):1092

    Article  Google Scholar 

  25. Sabarinathan D, Vanisree AJ (2011) Naringenin, a flavanone alters the tumorigenic features of C6 glioma cells. Biomed Prev Nutr 1(1):19–24

    Article  Google Scholar 

  26. Sabarinathan D, Mahalakshmi P, Vanisree AJ (2010) Naringenin promote apoptosis in cerebrally implanted C6 glioma cells. Mol Cell Biochem 345(1):215–222

    Article  CAS  Google Scholar 

  27. Chen K-Y, Chiu C-H, Wang L-C (2017) Anti-apoptotic effects of Sonic hedgehog signalling through oxidative stress reduction in astrocytes co-cultured with excretory-secretory products of larval Angiostrongylus cantonensis. Sci Rep 7(1):1–10

    Article  Google Scholar 

  28. Zhang RY et al (2018) Sonic hedgehog signaling regulates hypoxia/reoxygenation-induced H9C2 myocardial cell apoptosis. Exp Ther Med 16(5):4193–4200

    PubMed  PubMed Central  Google Scholar 

  29. Zhu S-L et al (2015) Sonic hedgehog signalling pathway regulates apoptosis through Smo protein in human umbilical vein endothelial cells. Rheumatology 54(6):1093–1102

    Article  CAS  Google Scholar 

  30. Bigelow RL et al (2004) Transcriptional regulation of bcl-2 mediated by the sonic hedgehog signaling pathway through gli-1. J Biol Chem 279(2):1197–1205

    Article  CAS  Google Scholar 

  31. Aroui S et al (2016) Naringin inhibits the invasion and migration of human glioblastoma cell via downregulation of MMP-2 and MMP-9 expression and inactivation of p38 signaling pathway. Tumor Biol 37(3):3831–3839

    Article  CAS  Google Scholar 

  32. Doheny D et al (2020) Hedgehog signaling and truncated GLI1 in cancer. Cells 9(9):2114

    Article  CAS  Google Scholar 

  33. Clement V et al (2007) HEDGEHOG-GLI1 signaling regulates human glioma growth, cancer stem cell self-renewal, and tumorigenicity. Curr Biol 17(2):165–172

    Article  CAS  Google Scholar 

  34. Skoda AM et al (2018) The role of the Hedgehog signaling pathway in cancer: a comprehensive review. Bosn J Basic Med Sci 18(1):8

    Article  CAS  Google Scholar 

  35. Wang K et al (2010) Sonic Hedgehog/GLI1 signaling pathway inhibition restricts cell migration and invasion in human gliomas. Neurol Res 32(9):975–980

    Article  Google Scholar 

  36. Bensalma S et al (2019) PKA at a cross-road of signaling pathways involved in the regulation of glioblastoma migration and invasion by the neuropeptides VIP and PACAP. Cancers 11(1):123

    Article  CAS  Google Scholar 

  37. Tayyab M et al (2019) Antidepressant and neuroprotective effects of naringenin via sonic hedgehog-GLI1 cell signaling pathway in a rat model of chronic unpredictable mild stress. NeuroMol Med 21(3):250–261

    Article  CAS  Google Scholar 

Download references

Funding

This research received a grant from Neuroscience Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran (Grant Number: 99000413). This study was approved by the Ethics Committee of Kerman University of Medical Sciences (IR.KMU.REC.1400.030).

Author information

Authors and Affiliations

  1. Physiology Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

    Marzieh Lotfian Sargazi

  2. Department of Pharmacology, School of Medicine, Semnan University of Medical Sciences, Semnan, Iran

    Kobra Bahrampour Juybari

  3. Herbal and Traditional Medicines Research Center, Kerman University of Medical Sciences, Kerman, Iran

    Mojdeh Esmaeili Tarzi & Mitra Mehrabani

  4. Pharmaceutical Sciences and Cosmetic Products Research Center, Kerman University of Medical Sciences, Kerman, Iran

    Arian Amirkhosravi

  5. Student Research Committee, Kerman University of Medical Sciences, Kerman, Iran

    Mohammad Hadi Nematollahi

  6. School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran

    Solmaz Mirzamohammdi

  7. Neuroscience Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran

    Mehrzad Mehrbani & Mehrnaz Mehrabani

Authors
  1. Marzieh Lotfian Sargazi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kobra Bahrampour Juybari
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mojdeh Esmaeili Tarzi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Arian Amirkhosravi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mohammad Hadi Nematollahi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Solmaz Mirzamohammdi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mehrzad Mehrbani
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Mehrnaz Mehrabani
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Mitra Mehrabani
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

MLS, MET, AA and SM carried out the experiment. KBJ and Mohammad HN wrote the manuscript and analysis data. MM and MM were involved in planning and supervised the work. MM helped shape the research and designed the figures.

Corresponding authors

Correspondence to Mehrnaz Mehrabani or Mitra Mehrabani.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Consent for participate

All authors have materially participated in the research and manuscript preparation.

Consent for publication

Consent for publication submission is approved by all authors.

Additional information

Publisher's Note

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

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sargazi, M.L., Juybari, K.B., Tarzi, M.E. et al. Naringenin attenuates cell viability and migration of C6 glioblastoma cell line: a possible role of hedgehog signaling pathway. Mol Biol Rep 48, 6413–6421 (2021). https://doi.org/10.1007/s11033-021-06641-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-021-06641-1

Keywords

Search

Navigation