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. 2016 Dec 27;7(52):87031-87036.
doi: 10.18632/oncotarget.13497.

The disintegrin echistatin in combination with doxorubicin targets high-metastatic human osteosarcoma overexpressing ανβ3 integrin in chick embryo and nude mouse models

Yasunori Tome  1   2   3 , Hiroaki Kimura  1   4 , Naotoshi Sugimoto  5 , Hiroyuki Tsuchiya  4 , Fuminori Kanaya  3 , Michael Bouvet  2 , Robert M Hoffman  1   2
Affiliations

The disintegrin echistatin in combination with doxorubicin targets high-metastatic human osteosarcoma overexpressing ανβ3 integrin in chick embryo and nude mouse models

Yasunori Tome et al. Oncotarget. .

Abstract

Echistatin, a cyclic RGD peptide, which is an antagonist of αvβ3 integrin (disintegrin), inhibited human osteosarcoma in the chick chorioallontoic membrane (CAM) model and tumor growth and pulmonary metastases in a nude mouse orthotopic model. A high-metastatic variant of human osteosarcoma, 143B-LM4, overexpressing αvβ3 integrin was used. Tumor angiogenesis by high-metastatic variant 143B-LM4 cells in the CAM was significantly inhibited by echistatin (P<0.05) as was overall growth. A doxorubicin (DOX)-echistatin combination inhibited orthotopic tumor growth compared to untreated control (P<0.01) or DOX alone (P<0.05) in nude mice. Tumor-bearing mice treated with the DOX-echistatin combination survived longer than those treated with DOX alone or control PBS (P<0.01 and P<0.01, respectively). Echistatin also inhibited experimental lung metastasis of 143B-LM4 cells in nude mice. These results suggest that DOX in combination with a disintegrin has potential to treat osteosarcoma and that αvβ3 integrin may be a target for osteosarcoma.

Keywords: echistatin; green fluorescent protein; nude mice; orthotopic; osteosarcoma; red fluorescent protein; αvβ3 integrin.

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Conflict of interest statement

CONFLICTS OF INTEREST

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1. Dual-color 143B-LM4 human osteosarcoma cells expressing GFP in the nucleus and RFP in the cytoplasm in vitro
The FV1000 confocal microscope (Olympus, Tokyo, Japan) was used to obtain images (20X).
Figure 2
Figure 2. Efficacy of echistatin on tumor-induced angiogenesis and tumor growth in the chick chorioallantoic membrane (CAM)
A. 143B-LM4-GEP-REP tumor fragments which grew on the CAM for 7 days were resected and implanted on another CAM which was treated with echistatin. Angiogenesis of the 143B-LM4 tumor was inhibited by echistatin compared with control (PBS). Tumor regression was also observed after treatment with echistatin. Bar: 2 mm. B. Quantitation of vessels vascularizing the tumor. Echistatin significantly decreased vascularization of the 143B tumor on the CAM (P<0.05). The OV100 variable-magnification fluorescence imager (Olympus, Tokyo, Japan) was used to obtain images. Error bars indicated SD.
Figure 3
Figure 3. Efficacy of echistatin in combination with DOX on the 143B-LM4 orthotopic model in nude mice
A. Efficacy of echistatin on metastasis in the 143B-LM4 orthotopic model in nude mice four weeks after transplantation in the tibia. The OV100 was used to obtain fluorescences images. Bar: 10 mm. B. Tumor volume four weeks after transplantation in the tibia. C. Kaplan-Meyer survival curves with the DOX-echistatin combination compared with control (PBS) or DOX-alone. P<0.01 for the DOX-echistatin combination on survival versus DOX- or PBS-treated animals.
Figure 4
Figure 4. Echistatin decreases the number of experimental lung metastases in 143B-LM4 cells
A. Fluorescence imaging of efficacy of echistatin on experimental metastasis of 143B-LM4-GFP-REP cells in nude mice one week after injection. The OV100 was used for fluorescence imaging. Bar: 2 mm. B. Quantitation of experimental lung metastases formed by 143B-LM4 cells. Echistatin decreased the number of experimental lung metastases in 143B-LM4 cells compared to untreated controls (P<0.01). Error bars indicate SD.
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