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Cancer Cell International
Published in: Cancer Cell International 1/2017

Open Access 01-12-2017 | Primary Research

Combined EphB2 receptor knockdown with radiation decreases cell viability and invasion in medulloblastoma

Authors: Shilpa Bhatia, Kellen Hirsch, Sanjana Bukkapatnam, Nimrah A. Baig, Ayman Oweida, Anastacia Griego, Dylan Calame, Jaspreet Sharma, Andrew Donson, Nicholas Foreman, Christopher Albanese, Sujatha Venkataraman, Rajeev Vibhakar, Sana D. Karam

Published in: Cancer Cell International | Issue 1/2017

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Abstract

Background

Medulloblastoma is one of the most common types of pediatric brain tumor characterized by the subpopulation of cells that exhibit high invasive potential and radioresistant properties. In addition, dysregulated function and signaling by Eph family of receptors have been shown to impart pro-tumorigenic characteristics in this brain malignancy. In the current study, we investigated whether EphB2 knockdown in combination with radiation can alter invasiveness and decrease medulloblastoma tumor growth or viability in vitro.

Methods

The expression of EphB2 receptor was analyzed by immunohistochemistry and Western blotting. Microarray analysis and mRNA analysis was performed on medulloblastoma patient datasets and compared to the normal cerebellum. The radiosensitization effect following EphB2 knockdown was determined by clonogenic assay in human medulloblastoma cells. Effects of EphB2-siRNA in absence or presence of radiation on cell cycle distribution, cell viability, and invasion were analyzed by flow cytometry, MTT assay, trypan blue exclusion assay, xcelligence system, and Western blotting.

Results

We observed that EphB2 is expressed in both medulloblastoma cell lines and patient samples and its downregulation sensitized these cells to radiation as evident by decreased clonogenic survival fractions. EphB2 expression was also high across different medulloblastoma subgroups compared to normal cerebellum. The radiosensitization effect observed following EphB2 knockdown was in part mediated by enhanced G2/M cell cycle arrest. We also found that the combined approach of EphB2 knockdown and radiation exposure significantly reduced overall cell viability in medulloblastoma cells compared to control groups. Similar results were obtained in the xcelligence-based invasion assay. Western blot analysis also demonstrated changes in the protein expression of cell proliferation, cell survival, and invasion molecules in the combination group versus others.

Conclusions

Overall, our findings indicate that specific targeting of EphB2 receptor in combination with radiation may serve as an effective therapeutic strategy in medulloblastoma. Future studies are warranted to test the efficacy of this approach in in vivo preclinical models.
Appendix
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Metadata
Title
Combined EphB2 receptor knockdown with radiation decreases cell viability and invasion in medulloblastoma
Authors
Shilpa Bhatia
Kellen Hirsch
Sanjana Bukkapatnam
Nimrah A. Baig
Ayman Oweida
Anastacia Griego
Dylan Calame
Jaspreet Sharma
Andrew Donson
Nicholas Foreman
Christopher Albanese
Sujatha Venkataraman
Rajeev Vibhakar
Sana D. Karam
Publication date
01-12-2017
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2017
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/s12935-017-0409-7

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