Title:Dyngo-4a Induces Neuroblastoma Cell Differentiation Through The AKT
and ERK1/2 Pathway
Volume: 22
Issue: 10
Author(s): Jinxi Huang*, Yi Zhou, Si Zeng, Jihong Xu, Lilian Liu, John Grothusen and Renyu Liu*
Affiliation:
- Department of General Surgery, The Affiliated Cancer Hospital of Zhengzhou University, Zhengzhou, Henan, China
- Department of Anesthesiology and Critical Care, Perelman School of Medicine at the University of Pennsylvania, 336
John Morgan Building, 3620 Hamilton Walk, PA 19104, Philadelphia, USA
- Department of Anesthesiology and Critical Care, Perelman School of Medicine at the University of Pennsylvania, 336
John Morgan Building, 3620 Hamilton Walk, PA 19104, Philadelphia, USA
Keywords:
Dyngo-4a, cell proliferation, retinoic acid, pathways, AKT, ERK1/2 pathway.
Abstract:
Aim: The aim of the study is to check whether dyngo-4a can inhibit neuroblastoma (NB)
proliferation and induce NB cell differentiation
Background: Dynamin plays a role in regulating neurotransmission, signaling pathways, nutrient uptake,
and pathogen infection, enhancing cell proliferation, tumor invasion, and metastasis. Studies
have reported that dyngo-4a, a dynamin inhibitor, can be used to identify potential biomarkers and
promising novel therapeutic targets for cancer treatment.
Objective: To our knowledge, no published reports are showing that dynamin inhibitors can reduce
NB cell proliferation and induce differentiation. In this study, we report that dyngo-4a can inhibit NB
proliferation and induce NB cell differentiation.
Methods: In this study, mouse neuroblastoma (Neuro-2a) cells were cultured in the presence or absence
of dyngo-4a or retinoic acid (RA), or in the presence of both dyngo-4a and RA, or in the presence
of sequential administration of dyngo-4a and RA to compare the effects on the inhibition of cell
proliferation and effects on neuroblastoma cell differentiation induction. The neural cell markers, Nestin
and Tuj 1 (Neuron-specific class III beta-tubulin), were used to demonstrate that the differentiated
cells have neuronal cell features. The phosphorylation of Protein Kinase B (AKT), extracellular signalregulated
kinases1/2 (ERK1/2), and epidermal growth factor receptor (EGFR) were determined to examine
the potential mechanisms of induced differentiation.
Results: Dyngo-4a or RA or dyngo-4a with subsequent RA administration induced Neuro-2a cell differentiation.
However, RA with subsequent dyngo-4a administration results in almost total death of
the Neuro-2a cells. The differentiation rate induced by dyngo-4a was significantly higher than the rate
by RA treatment (72.5 ± 1.4% vs. 52.9 ± 3.1% with neuron features, P<0.05; 39.0 ± 0.8% vs. 29.9 ±
1.8% for axons under light microscopy, p<0.05). The differentiation rate of cells treated with dyngo-4a
first, followed by RA, was greater than when they were added together (74.8 ± 3.8% vs. 10.6 ± 3.6%;
45.5 ± 1.6% vs. 12.4 ± 0.6%, p<0.01). Co-administration of dyngo-4a and RA at the same time diminished
differentiation efficacy significantly. Dyngo-4a induced Neuro-2a cell differentiation and increased
Tuj-1 positive staining by the 6th day post- treatment. Dyngo-4a also inhibited Neuro-2a cell
proliferation in a dose-dependent manner. Regarding the mechanism, dyngo-4a treatment showed a
significant increase in p-AKT and p-ERK1/2 but not in p-EGFR.
Conclusion: At a level comparable to RA, dynamin inhibition with dyngo-4a lowers proliferation and
causes differentiation of Neuro-2a mouse NB cells in vitro. The AKT pathway is activated by dynago-
4a, which results in differentiation. The combination of RA with dynago-4a reduces the efficiency of
differentiation. The application of dynago-4a followed by RA, on the other hand, enhances the differentiating
effect, implying alternative mechanistic roles in the process.
