5 June 2012 Effect of antiangiogenic therapy on luciferase activity in a cytomegalovirus- or HSP70-promoter-transfected M21 tumor model
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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.
© 2012 Society of Photo-Optical Instrumentation Engineers (SPIE)
Walter Hundt, Walter Hundt, Christian Schink, Christian Schink, Silke Steinbach, Silke Steinbach, Caitlin E. O’Connell-Rodwell, Caitlin E. O’Connell-Rodwell, Andreas Kiessling, Andreas Kiessling, Damiano Librizzi, Damiano Librizzi, Mykhaylo Burbelko, Mykhaylo Burbelko, Samira Guccione, Samira Guccione, } "Effect of antiangiogenic therapy on luciferase activity in a cytomegalovirus- or HSP70-promoter-transfected M21 tumor model," Journal of Biomedical Optics 17(6), 065001 (5 June 2012). https://doi.org/10.1117/1.JBO.17.6.065001 . Submission: Received: 23 February 2012; Accepted: 24 April 2012
Received: 23 February 2012; Accepted: 24 April 2012; Published: 5 June 2012
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