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

Mol Biol Rep. 2021 Sep;48(9):6413-6421. doi: 10.1007/s11033-021-06641-1. Epub 2021 Aug 24.

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.

Keywords: C6; Glioblastoma; Hedgehog signaling pathway; Migration; Naringenin.

MeSH terms

  • 3T3 Cells
  • Animals
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / drug effects
  • Brain Neoplasms / metabolism*
  • Brain Neoplasms / pathology
  • Cell Line, Tumor
  • Cell Movement / drug effects*
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Flavanones / pharmacology*
  • Glioblastoma / metabolism*
  • Glioblastoma / pathology
  • Mice
  • Rats
  • Repressor Proteins / metabolism*
  • Signal Transduction / drug effects*
  • Smoothened Receptor / metabolism*
  • Zinc Finger Protein GLI1 / metabolism*

Substances

  • Antineoplastic Agents
  • Flavanones
  • Gli1 protein, rat
  • Repressor Proteins
  • Smo protein, rat
  • Smoothened Receptor
  • SuFu protein, rat
  • Zinc Finger Protein GLI1
  • naringenin