April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Inhibition of Retinoblastoma in vitro and in vivo With Conditionally Replicating Oncolytic Adenovirus H101
Author Affiliations & Notes
  • X. Song
    Ophthalmology, Shanghai 9th People's Hospital, Shanghai, China
  • X. Fan
    Ophthalmology, Shanghai 9th People's Hospital, Shanghai, China
  • R. Jia
    Ophthalmology, Shanghai 9th People's Hospital, Shanghai, China
  • Y. Zhou
    Ophthalmology, Shanghai 9th People's Hospital, Shanghai, China
  • X. Xu
    Ophthalmology, Shanghai 9th People's Hospital, Shanghai, China
  • H. Wang
    Department of Biochemistry and Molecular Biology, Shanghai Jiaotong University School of Medicine, Shanghai, China
  • S. Ge
    Department of Biochemistry and Molecular Biology, Shanghai Jiaotong University School of Medicine, Shanghai, China
  • Footnotes
    Commercial Relationships  X. Song, None; X. Fan, None; R. Jia, None; Y. Zhou, None; X. Xu, None; H. Wang, None; S. Ge, None.
  • Footnotes
    Support  Shanghai Leading Academic Discipline Project S30205 ; Science and Technology Commission of Shanghai 08410702300
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 1693. doi:
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      X. Song, X. Fan, R. Jia, Y. Zhou, X. Xu, H. Wang, S. Ge; Inhibition of Retinoblastoma in vitro and in vivo With Conditionally Replicating Oncolytic Adenovirus H101. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1693.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To determine the functional effects of oncolytic adenovirus H101 on retinoblastoma in vitro and in vivo.

Methods: : The expression of Coxsackie-adenovirus receptor (CAR) in the human retinoblastoma cell line HXO-RB44 was determined by reverse transcription polymerase chain reaction (RT-PCR). Retinoblastoma cells were incubated with green fluorescent protein (GFP)-labeled adenovirus (AdGFP) and appropriate multiplicity of infection (MOI) was determined using flow cytometry. Cell viability of HXO-RB44 treated with H101 or AdGFP was measured using a CCK8-based procedure. Viral proliferation was measured through endpoint dilution titration on 293 human embryonic kidney cells and real-time PCR. Cell cycle and apoptosis were analyzed by flow cytometry. NOD-SCID mice bearing retinoblastoma xenografts were treated with intratumoral injection of H101, AdGFP, or phosphate-buffered saline. Tumor volume and survival time were recorded. Immunohistochemistry for adenoviral fiber protein in paraffin-embedded tissue sections of retinoblastoma xenografts was performed to evaluate H101 virus replication in vivo.

Results: : HXO-RB44 cells extensively expressed CAR and were sensitive to adenoviral infection. HXO-RB44 cells treated with H101 had dramatically reduced cell viability compared with AdGFP-treated cells (P<0.01). Abundant replication of H101 was observed in HXO-RB44 cells, resulting in G2/M phase arrest, but there was no apoptosis. Tumor-bearing mice treated with H101 had significantly reduced tumor volume (68%) and significantly prolonged survival time (170%) compared with controls (both P<0.01). Immunohistochemical examination revealed widespread replication of H101 within the tumor.

Conclusions: : These results suggest that oncolytic adenovirus H101 effectively inhibits retinoblastoma cells in vitro and in vivo, and may be useful for the clinical treatment of retinoblastoma.

Keywords: retinoblastoma • gene transfer/gene therapy • adenovirus 
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