Related Experiment Video
Updated: Feb 3, 2026
An Improved Protocol to Purify and Directly Mono-Biotinylate Recombinant BDNF in a Tube for Cellular Trafficking Studies in Neurons
Published on: July 11, 2020
P2X4R silence suppresses glioma cell growth through BDNF/TrkB/ATF4 signaling pathway
Jun-Feng Huo1, Xiao-Bing Chen1
1Second Ward, Department of Neurosurgery, Huaihe Hospital of Henan University, Kaifeng, China.
Abstract:
Purinergic receptor P2X 4 (P2X4R), a member of purinergic channels family and a subtype of ionotropic adenosine triphosphate receptors, plays a critical role in tumorigenesis. Evidence suggested that P2X4R is expressed in rat C6 glioma model, however, its role and the underlying mechanism of action are still unclear in human glioblastoma multiforme (GBM). In the current study, our aim is to examine the function and the molecular basis of P2X4R in GBM. We first observed that GBM cells, U251, T98, U87, U373, and A172 were all high expressed P2X4R, when compared with the normal human astrocytes (NHA) cells. To gain the function of P2X4R, P2X4R silence cells were constructed by transfection with P2X4R small interfering RNA (siRNA). We found that P2X4R deletion impeded T98 and U87 cell viability and proliferation, and further studies indicated that cell apoptosis and caspase-3 activity was increased in T98 and U87 cell transfected with P2X4R siRNA. Subsequently, we confirmed that P2X4R silence suppressed brain-derived neurotrophic factor (BDNF), Trk receptor tyrosine kinases (TrkB), and activating transcription factor 4 (ATF4) expression in T98 and U87 cells. And P2X4R siRNA-induced ATF4-expression inhibition dependent on BDNF/TrkB signaling pathway. The impact of P2X4R silence on T98 and U87 cell growth and apoptosis was reversed by ATF4 overexpression. In summary, this study provides the first evidence that P2X4R plays important roles in GBM cell growth and apoptosis.
Insights
Purinergic receptor P2X4 (P2X4R) is highly expressed in glioblastoma multiforme (GBM) cells. Silencing P2X4R inhibits GBM cell growth and induces apoptosis via the BDNF/TrkB/ATF4 pathway.
Area of Science:
- Neuroscience
- Molecular Biology
- Oncology
Background:
- Purinergic receptor P2X 4 (P2X4R), an ionotropic ATP receptor, is implicated in tumorigenesis.
- While P2X4R expression is noted in rat glioma models, its specific role in human glioblastoma multiforme (GBM) remains unclear.
Purpose of the Study:
- To investigate the function and molecular mechanisms of P2X4R in human GBM.
- To determine the impact of P2X4R on GBM cell proliferation, apoptosis, and related signaling pathways.
Main Methods:
- Assessed P2X4R expression in human GBM cell lines (U251, T98, U87, U373, A172) and normal human astrocytes (NHA).
- Utilized small interfering RNA (siRNA) to silence P2X4R in T98 and U87 GBM cells.
- Analyzed cell viability, proliferation, apoptosis, caspase-3 activity, and expression of BDNF, TrkB, and ATF4.
- Investigated the role of the BDNF/TrkB/ATF4 pathway and ATF4 overexpression in P2X4R-silenced cells.
Main Results:
- GBM cells exhibited significantly higher P2X4R expression compared to NHA cells.
- P2X4R silencing in T98 and U87 cells reduced cell viability and proliferation.
- P2X4R knockdown led to increased apoptosis and caspase-3 activity in GBM cells.
- Silencing P2X4R suppressed BDNF, TrkB, and ATF4 expression, with ATF4 inhibition dependent on the BDNF/TrkB pathway.
- Overexpression of ATF4 reversed the effects of P2X4R silencing on GBM cell growth and apoptosis.
Conclusions:
- P2X4R plays a significant role in promoting GBM cell growth and inhibiting apoptosis.
- The P2X4R signaling pathway involves BDNF, TrkB, and ATF4 in the context of GBM.
- Targeting P2X4R may represent a potential therapeutic strategy for glioblastoma multiforme.
Related Concept Videos
03:14Notch Signaling Pathway
The Notch gene came into the limelight in 1914 after the discovery that its mutation in Drosophila melanogaster leads to a serrated (or "notched") wing margin phenotype. It was not...
02:33Hedgehog Signaling Pathway
01:19Interactions Between Signaling Pathways
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
01:41Non-Canonical Wnt Signaling Pathways
02:54Canonical Wnt Signaling Pathway
02:26NF-κB-dependent Signaling Pathway
NF-κB-dependent Signaling Mechanism
The...