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Published on: February 22, 2015
PTBP1 knockdown promotes neural differentiation of glioblastoma cells through UNC5B receptor
Kankai Wang1,2, Sishi Pan1,2, Peiqi Zhao1,2
1Zhejiang Provincial Key Laboratory of Aging and Neurological Disorder Research, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, China.
Abstract:
Rationale: Cell reprogramming technology is utilized to prevent cancer progression by transforming cells into terminally differentiated, non-proliferating states. Polypyrimidine tract binding protein 1 (PTBP1) is an RNA binding protein required for the growth of neurons and may directly transform multiple normal human cells into functioning neurons in vitro and in vivo when expressed at low levels. As a result, we identified it as a key to inhibiting cancer cell proliferation by boosting glioblastoma cell neural differentiation. Methods: Immunocytofluorescence (ICF) targeting TUJ1, MAP2, KI67, and EdU were utilized to evaluate glioblastoma cell reprogramming under PTBP1 knockdown or other conditions. PTBP1 and other target genes were detected using Western blotting and qRT-PCR. Activating protein phosphatase 2A (PP2A) and RhoA were detected using specific kits. CCK8 assays were employed to detect cell viability. Bioluminescence, immunohistofluorescence (IHF), and Kaplan-Meier survival analyses were utilized to demonstrate the in vivo reprogramming efficiency of PTBP1 knockdown in U87 murine glioblastoma model. In this study, RNA-seq technology was used to examine the intrinsic pathway. Results: The expression of TUJ1 and MAP2 neural markers, as well as the absence of KI67 and EdU proliferative markers in U251, U87, and KNS89 cells, indicated that glioblastoma cell reprogramming was successful. In vivo, U87 growth generated xenografts was substantially shrank due to PTBP1 knockdown induced neural differentiation, and these tumor-bearing mice had a prolonged survival time. Following RNA-seq, ten potential downstream genes were eliminated. Lentiviral interference and inhibitors blocking tests demonstrated that UNC5B receptor and its downstream signaling were essential in the neural differentiation process mediated by PTBP1 knockdown in glioblastoma cells. Conclusions: Our results indicate that PTBP1 knockdown promotes neural differentiation of glioblastoma cells via UNC5B receptor, consequently suppressing cancer cell proliferation in vitro and in vivo, providing a promising and feasible approach for glioblastoma treatment.
Insights
Knocking down Polypyrimidine tract binding protein 1 (PTBP1) induces neural differentiation in glioblastoma cells, inhibiting proliferation. This approach, targeting the UNC5B receptor, offers a promising new treatment strategy for glioblastoma.
Area of Science:
- Neuroscience
- Oncology
- Molecular Biology
Background:
- Cell reprogramming offers a strategy to halt cancer progression by inducing terminal differentiation.
- Polypyrimidine tract binding protein 1 (PTBP1) is crucial for neuronal development and can induce neural differentiation in non-neuronal cells.
Purpose of the Study:
- To investigate the potential of PTBP1 as a target for inhibiting glioblastoma (GBM) cell proliferation through induced neural differentiation.
- To elucidate the molecular mechanisms underlying PTBP1-mediated neural differentiation in GBM cells.
Main Methods:
- Glioblastoma cell reprogramming was assessed using immunocytochemistry (TUJ1, MAP2, KI67, EdU) and molecular analyses (Western blot, qRT-PCR).
- Cell viability was measured by CCK8 assays.
- In vivo efficacy was evaluated in a U87 murine glioblastoma model using bioluminescence, IHF, and survival analysis. RNA-sequencing identified downstream pathways.
Main Results:
- PTBP1 knockdown successfully induced neural differentiation markers (TUJ1, MAP2) and suppressed proliferation markers (KI67, EdU) in U251, U87, and KNS89 cells.
- In vivo, PTBP1 knockdown significantly reduced U87 xenograft growth and prolonged survival in tumor-bearing mice.
- RNA-seq and subsequent functional studies identified the UNC5B receptor and its downstream signaling as critical mediators of PTBP1 knockdown-induced neural differentiation.
Conclusions:
- PTBP1 knockdown promotes glioblastoma cell neural differentiation via the UNC5B receptor pathway.
- This process effectively suppresses glioblastoma cell proliferation both in vitro and in vivo.
- Targeting PTBP1 represents a novel and viable therapeutic strategy for glioblastoma treatment.
