Inhibitory effects of subcutaneous tumors in nude mice mediated by low‑frequency ultrasound and microbubbles

Shen,Zhi‑Yong; Shen,E.; Diao,Xue‑Hong; Bai,Wen‑Kun; Zeng,Min‑Xia; Luan,Yan Yan; Nan,Shu‑Liang; Lin,Yan‑Duan; Wei,Cong; Chen,Li; Sun,Di; Hu,Bing

doi:10.3892/ol.2014.1934

Inhibitory effects of subcutaneous tumors in nude mice mediated by low‑frequency ultrasound and microbubbles

Authors:
- Zhi‑Yong Shen
- E. Shen
- Xue‑Hong Diao
- Wen‑Kun Bai
- Min‑Xia Zeng
- Yan Yan Luan
- Shu‑Liang Nan
- Yan‑Duan Lin
- Cong Wei
- Li Chen
- Di Sun
- Bing Hu
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Affiliations: Department of Ultrasound in Medicine, Shanghai Institute of Ultrasound in Medicine, Sixth People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai Jiaotong University School of Medicine, Shanghai 200233, P.R. China
Published online on: March 4, 2014 https://doi.org/10.3892/ol.2014.1934
Pages: 1385-1390

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Abstract

The aim of the present study was to investigate the sonication effects of 21‑kHz ultrasound (US) with microbubbles (MBs) on the subcutaneous prostate tumors of nude mice. In total, 15 tumor‑bearing nude mice were divided into three groups: The control group, the low‑frequency US group and the US+MB group. The MBs used were from US contrast agent SonoVue. The parameters of the US were as follows: 21 kHz, 26 mW/cm2 and a 40% duty cycle (2 sec on, 3 sec off) for 3 min, once every other day for 2 weeks. Color Doppler flow imaging, hematoxylin and eosin (HE) staining, immunoblotting and transmission electron microscopy (TEM) were used to evaluate the results. Following 2 weeks of treatment, the blood flow signal disappeared in the US+MB group only, and the tumor size was smaller when compared with the control and US groups. For the immunoblotting, the intensity of cyclooxygenase‑2 and vascular endothelial growth factor in the US+MB group was lower compared with the other two groups. Tumor necrosis was present and the nucleus disappeared upon HE staining in the US+MB group. Upon TEM analysis, increased cytoplasmic vacuolation and dilatation of the perinuclear cisternae of the tumor cells were found in the US+MB group. In the control and US groups, the tumors had intact vascular endothelia and vessel lumens. However, lumen occlusion of the vessels was observed in the US+MB group. In conclusion, 21‑kHz low‑intensity US with MBs may result in vessel occlusion and growth inhibitory effects in the subcutaneous tumors of nude mice.

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