Korean J Radiol.  2015 Aug;16(4):776-782. 10.3348/kjr.2015.16.4.776.

Combined Radiofrequency Ablation and Double Anti-Angiogenic Protein Therapy to Increase Coagulation Efficacy: An Experimental Study in a Murine Renal Carcinoma Model

• Jun HY
• Ryu JH
• Byun SJ
• Jeong CW
• Kim TH
• Lee YH
• Yoon KH

• KMID: 2155550
• DOI: http://doi.org/10.3348/kjr.2015.16.4.776

Abstract

OBJECTIVE
To evaluate whether suppression of tumor microvasculature by double anti-angiogenic protein (DAAP) treatment could increase the extent of radiofrequency ablation (RFA)-induced coagulation in a murine renal cell carcinoma model.
MATERIALS AND METHODS
Renal cell carcinoma cell lines were implanted subcutaneously into 10 nude mice. Four mice received adenoviral DAAP treatment and 6 mice received sterile 0.9% saline solution as DAAP-untreated group. The effect of DAAP was evaluated according to the vascularity by contrast-enhanced ultrasound (CEUS) using microbubbles. Four DAAP-treated mice and 4 DAAP-untreated mice were then treated with RFA, resulting in 3 groups: no-therapy (n = 2), RFA only (n = 4), and RFA combined with DAAP treatment (n = 4). Immediately after RFA, the size of coagulation necrosis and mitochondrial enzyme activity were compared between the groups using analysis of variance (ANOVA) and post hoc test.
RESULTS
The contrast enhancement ratio for tumor vascularization on CEUS was significantly lower in the DAAP treated group than in DAAP-untreated group (30.2 +/- 9.9% vs. 77.4 +/- 17.3%; p = 0.021). After RFA, the mean coagulation diameter was 0 mm for no-therapy group, 6.7 +/- 0.7 mm for the RFA only group and 8.5 +/- 0.4 mm for the RFA with DAAP group (ANOVA, p < 0.001). The area of viable mitochondria within the tumor was 27.9 +/- 3.9% in no-therapy group, 10.3 +/- 4.5% in the RFA only group, and 2.1 +/- 0.7% in the RFA with DAAP group (ANOVA, p < 0.001).
CONCLUSION
Our results suggest the potential value of combining RFA with anti-angiogenic therapy.

Keyword

Radiofrequency ablation; Angiogenesis; Anti-angiogenic protein; Renal cell carcinoma; Mice model

MeSH Terms

Adenoviridae
Angiogenic Proteins/*antagonists & inhibitors
Animals
Carcinoma, Renal Cell/blood supply/surgery/*therapy
Catheter Ablation/*methods
Combined Modality Therapy
Contrast Media
Kidney Neoplasms/blood supply/surgery/*therapy
Male
Mice
Mice, Nude
Microbubbles
Neovascularization, Pathologic/surgery/*therapy/ultrasonography
Recombinant Proteins
Angiogenic Proteins
Contrast Media
Recombinant Proteins

Figure

• 

  Fig. 1 Comparison of contrast-enhanced ultrasound images of tumors after injection of microbubbles. A, B. Images of untreated tumor show strong enhancement in intratumor area (yellow circle) after injection of microbubbles. C, D. In contrast, treated tumor with double anti-angiogenic protein (DAAP) shows similar contrast enhancement after injection of microbubbles.

• 

  Fig. 2 Graph demonstrating contrast enhancement ratios (CER). CER after injection of microbubbles was significantly different between untreated tumor and double anti-angiogenic protein (DAAP)-treated tumor. *p < 0.05.

• 

  Fig. 3 Assessment for mitochondrial viable area. Cross sections of three gross pathologic specimens stained with 2,3,5-triphenyl-2H-tetrazolium chloride. Mitochondrial activity is shown in red. Nonviable ablated tumor remains white (arrows). Mitochondrial activity is clearly seen in (A) untreated tumor but not in treated tumor with (B) radiofrequency ablation (RFA) only or with (C) RFA and double anti-angiogenic protein (DAAP). (D-F) Remaining viable mitochondria were overlaid as red color using MATLAB.

• 

  Fig. 4 Graph demonstrating area of viable mitochondria. Tumor treated with RFA and DAAP showed significantly smaller zone of mitochondrial activity than no-therapy group and RFA-only group. P values among 'no-therapy', 'RFA only', and 'RFA with DAAP' are indicated in graph (one-way ANOVA followed by post hoc Tukey's test). ANOVA = analysis of variance, DAAP = double anti-angiogenic protein, RFA = radiofrequency ablation

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