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Neuroblastoma Cell Death Induced by eEF1A2 Knockdown Is Possibly Mediated by the Inhibition of Akt and mTOR

Kawinthra Khwanraj1, Permphan Dharmasaroja1

  • 1Department of Anatomy, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.

International Journal of Hematology-Oncology and Stem Cell Research
|March 16, 2022
PubMed

Insights

Silencing eukaryotic translation elongation factor-1, alpha-2 (eEF1A2) reduced neuroblastoma cell viability. This inhibition occurred via the protein kinase B/mammalian target of the rapamycin (Akt/mTOR) pathway, suggesting eEF1A2 as a therapeutic target.

Area of Science:

  • Oncology
  • Molecular Biology
  • Cell Biology

Background:

  • Neuroblastoma exhibits constitutive activation of the prosurvival protein kinase B/mammalian target of the rapamycin (Akt/mTOR) pathway.
  • Eukaryotic translation elongation factor-1, alpha-2 (eEF1A2) protein activates the Akt/mTOR pathway, but its role in neuroblastoma is understudied.

Purpose of the Study:

  • To investigate the impact of eEF1A2 silencing on neuroblastoma cell viability.
  • To explore the associated signaling mechanisms, particularly the Akt/mTOR pathway.

Main Methods:

  • Human SH-SY5Y neuroblastoma cells were transfected with small interfering RNA (siRNA) targeting eEF1A2.
  • Cell viability was assessed using MTT assay.
  • mRNA and protein expression of key apoptosis and signaling pathway markers (p53, Bax, Bcl-2, caspase-3, PI3K, Akt, mTOR) were analyzed via qRT-PCR and Western blot.

Main Results:

  • eEF1A2 knockdown significantly decreased neuroblastoma cell viability.
  • While mRNA levels of p53, Bax/Bcl-2 ratio, and caspase-3 showed upregulated trends, they were not statistically significant.
  • eEF1A2 silencing notably inhibited the phosphorylation of Akt and mTOR, with slight alterations in PI3K, Akt2, mTORC1, and mTORC2 expression.

Conclusions:

  • eEF1A2 knockdown induces neuroblastoma cell death.
  • This effect is mediated, at least in part, by the inhibition of the Akt/mTOR signaling pathway.
  • eEF1A2 presents a potential molecular target for neuroblastoma therapeutic strategies.
Keywords:
AktEukaryotic translation elongation factor-1 alpha-2 (eEF1A2)NeuroblastomaPhosphoinositide 3-kinases (PI3K)SH-SY5Y cellsSmall interfering RNA (siRNA)mTORp53

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