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Published on: October 4, 2019
GliPR1 knockdown by RNA interference exerts anti-glioma effects in vitro and in vivo
Urban J Scheuring1, Steffi Ritter2, Daniel Martin2
1Department of Hematology/Oncology and Infectious Diseases, University Hospital, J.W. Goethe University, Theodor-Stern-Kai 7, 60590, Frankfurt am Main, Germany. u.scheuring@gmx.de.
Introduction:
In human glioblastomas, glioma pathogenesis-related protein1 (GliPR1) is overexpressed and appears to be an oncoprotein. We investigated whether GliPR1 knockdown in glioma cells by RNA interference exerts anti-glioma effects.
Methods:
Experiments used human glioblastoma cell lines transduced with GliPR1 shRNA (sh#301, sh#258). Transduction produced stringent doxycycline-dependent GliPR1 knockdown in clones (via lentiviral "all-in-one" TetOn-shRNA vector) or stable GliPR1 knockdown in polyclonal cells (via constitutive retroviral-shRNA vector). In vitro assessments included cellular proliferation and clonogenic survival. In vivo assessments in tumor-bearing nude mice included tumor growth and survival.
Results:
Using doxycycline-dependent GliPR1 knockdown, shGliPR1-transduced U87-MG clones demonstrated reductions in cellular proliferation in the presence versus absence of doxycycline. Using stable GliPR1 knockdown, polyclonal shGliPR1-transduced U87-MG, A172, and U343-MG cells consistently showed decreased clonogenic survival and induced apoptosis (higher proportion of early apoptotic cells) compared to control shLuc-transduced cells. In tumor-bearing nude mice, using doxycycline-dependent GliPR1 knockdown, subcutaneous and cranial transplantation of the U87-MG clone 980-5 (transduced with GliPR1 sh#301) resulted in reduced subcutaneous tumor volume and cerebral tumor area in doxycycline-treated mice versus those left untreated. Using stable GliPR1 knockdown, nude mice cranially transplanted with polyclonal U87-MG cells transduced with GliPR1 sh#258 had significantly prolonged survival compared to mice cranially transplanted with control shLuc-transduced cells (41 versus 26 days; P < 0.001).
Conclusion:
GliPR1 knockdown in glioma cells decreased cellular proliferation, decreased clonogenic survival, and induced apoptosis in vitro, and reduced glioblastoma tumor growth and prolonged survival in vivo. These findings support that GliPR1 may have potential value as a therapeutic target.
Insights
GliPR1 knockdown in glioma cells inhibits proliferation and survival. This study shows GliPR1 (glioma pathogenesis-related protein1) knockdown reduces glioblastoma growth and prolongs survival in vivo, suggesting it as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- RNA Interference
Background:
- Glioma pathogenesis-related protein1 (GliPR1) is overexpressed in human glioblastomas and functions as an oncoprotein.
- Investigating the anti-glioma effects of GliPR1 knockdown is crucial for developing targeted therapies.
Purpose of the Study:
- To determine if RNA interference-mediated knockdown of GliPR1 in glioma cells exerts anti-glioma effects.
- To evaluate the impact of GliPR1 knockdown on glioblastoma cell proliferation, survival, and tumor growth in vivo.
Main Methods:
- Human glioblastoma cell lines were transduced with GliPR1 shRNA (sh#301, sh#258) using lentiviral or retroviral vectors for doxycycline-dependent or stable knockdown, respectively.
- In vitro assessments included cellular proliferation and clonogenic survival assays.
- In vivo studies involved tumor-bearing nude mice, assessing tumor growth and animal survival.
Main Results:
- GliPR1 knockdown significantly reduced cellular proliferation and induced apoptosis in glioblastoma cell lines in vitro.
- In vivo, GliPR1 knockdown led to reduced subcutaneous tumor volume and cerebral tumor area.
- Stable GliPR1 knockdown significantly prolonged survival in tumor-bearing nude mice.
Conclusions:
- GliPR1 knockdown effectively inhibits glioblastoma cell proliferation and survival in vitro.
- GliPR1 knockdown demonstrates significant anti-tumor efficacy and improves survival in vivo.
- These findings highlight GliPR1 as a potential therapeutic target for glioblastoma treatment.
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