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Journal of Biomedical Optics

Effect of antiangiogenic therapy on luciferase activity in a cytomegalovirus- or HSP70-promoter-transfected M21 tumor model
Author(s): Walter Hundt; Samira Guccione; Andreas Kiessling; Mykhaylo Burbelko; Christian Schink; Silke Steinbach; Caitlin E. O'Connell-Rodwell; Damiano Librizzi
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Paper Abstract

We investigated the effect of targeted gene therapy on heat shock protein 70 expression (Hsp70) and protein production (HSP70) in a melanoma tumor model (M21; M21-L). M21 and M21-L cells transfected with a plasmid containing the Hsp70 (Hspa1b) or the cytomegalovirus (CMV) promoter and the luciferase reporter gene were injected into mice; the resulting tumors grew to a size of 650  mm3. Mice (five per group) were intravenously treated with an Arg-Gly-Asp peptide-nanoparticle/Raf-1 kinase inhibitor protein complex [RGD-NP/RAF(-)] or with a nanoparticle control. Bioluminescence imaging (IVIS®, Xenogen, USA) was performed at 12, 24, 48, and 72 h after the treatment cycle. Western blot analysis of HSP70 protein was performed to monitor protein expression. The size of the treated M21 tumors remained fairly constant (647.8±103.4  mm2 at the beginning versus 704.8±94.4  mm3 at the end of the experiment). The size of the M21-L tumors increased, similar to the untreated control tumors. Bioluminescent imaging demonstrated that when transcription was controlled by the CMV promoter, luciferase activity decreased to 17.9%±4.3% of baseline values in the treated M21 tumors. When transcription was controlled by the Hsp70 promoter, the highest luciferase activity (4.5±0.7-fold increase over base-line values) was seen 24 h after injection in the M21 tumors; however, no luciferase activity was seen in the M21-L tumors. In accordance with bioluminescent imaging, western blot analysis showed a peak in HSP70 production at 24 h after the injection of the RGD-NP/RAF(-) complex in the M21 tumors; however, no HSP70 protein induction was seen in the M21-L tumors. Thus, targeted antiangiogenic therapy can induce Hsp70 expression and HSP70 protein in melanoma tumors.

Paper Details

Date Published: 5 June 2012
PDF: 8 pages
J. Biomed. Opt. 17(6) 065001 doi: 10.1117/1.JBO.17.6.065001
Published in: Journal of Biomedical Optics Volume 17, Issue 6
Show Author Affiliations
Walter Hundt, Stanford Univ. School of Medicine (United States)
Samira Guccione, Stanford Univ. School of Medicine (United States)
Andreas Kiessling, Philipps-Univ. Marburg (Germany)
Mykhaylo Burbelko, Philipps-Univ. Marburg (Germany)
Christian Schink, Philipps-Univ. Marburg (Germany)
Silke Steinbach, Philipps-Univ. Marburg (Germany)
Caitlin E. O'Connell-Rodwell, Stanford Univ. School of Medicine United (United States)
Damiano Librizzi, Philipps-Univ. Marburg (Germany)


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