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Inhibitory effects of subcutaneous tumors in nude mice mediated by low-frequency ultrasound and microbubbles
Zhi-Yong Shen1, E Shen2, Xue-Hong Diao2
1Department 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 ; Department of Radiology, Nantong Tumor Hospital Affiliated to Nantong University, Nantong, Jiangsu 226361, P.R. China.
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.
Insights
Low-frequency ultrasound (US) combined with microbubbles (MBs) effectively reduced subcutaneous prostate tumor size in mice. This combination therapy led to tumor vessel occlusion and inhibited tumor growth by targeting key molecular pathways.
Area of Science:
- Oncology
- Biomedical Engineering
- Pharmacology
Background:
- Subcutaneous prostate tumors in nude mice are a common model for studying tumor growth and treatment efficacy.
- Ultrasound (US) and microbubbles (MBs) have emerged as potential therapeutic agents, but their combined effects on solid tumors require further investigation.
- Understanding the mechanisms of US-MB interaction is crucial for developing targeted cancer therapies.
Purpose of the Study:
- To investigate the effects of 21-kHz low-frequency ultrasound (US) combined with microbubbles (MBs) on subcutaneous prostate tumors in nude mice.
- To evaluate the impact of this combination therapy on tumor vascularization, growth, and cellular morphology.
- To explore the molecular changes associated with US-MB treatment, focusing on cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF).
Main Methods:
- 15 nude mice with subcutaneous prostate tumors were divided into three groups: control, US only, and US + MBs (SonoVue).
- Treatment involved 21-kHz US (26 mW/cm², 40% duty cycle) for 3 minutes every other day for 2 weeks.
- Evaluations included Color Doppler, hematoxylin and eosin (HE) staining, immunoblotting, and transmission electron microscopy (TEM).
Main Results:
- The US + MB group showed complete disappearance of blood flow signals and significantly smaller tumor size compared to control and US-only groups.
- Immunoblotting revealed lower levels of COX-2 and VEGF in the US + MB group.
- Histological analysis (HE staining) indicated tumor necrosis and nuclear disappearance in the US + MB group, while TEM showed cytoplasmic vacuolation and perinuclear cisterna dilatation, along with vessel lumen occlusion.
Conclusions:
- 21-kHz low-intensity ultrasound combined with microbubbles effectively inhibits subcutaneous tumor growth in mice.
- The observed anti-tumor effects are attributed to vascular occlusion within the tumor, leading to necrosis and reduced expression of key angiogenic factors.
- This combination therapy presents a promising approach for localized tumor treatment, warranting further investigation in clinical settings.
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