Department of Ultrasound, Southwest Hospital, Third Military Medical University (Army Medical University), 30 Gaotanyan Street, Shapingba District, Chongqing 400038, People's Republic of China.
Nanotechnology. 2020 Mar 27;31(24):245102. doi: 10.1088/1361-6528/ab7e73. Epub 2020 Mar 10.
To construct targeted nanobubbles carrying both small-molecule CXCR4 antagonist AMD070 and light-absorbing material indocyanine green (ICG), and to study their in vitro multimodal imaging, as well as their mechanism and efficacy of inhibition of breast cancer cell growth. Nanobubbles carrying AMD070 and ICG (ICG-TNBs) were constructed by carbodiimide reaction and mechanical oscillation. The physical characteristics and in vitro multimodal imaging were determined. The binding potential of ICG-TNBs to human breast cancer cells were observed by laser confocal microscopy. CCK-8 and flow cytometry were used to analyze the role of ICG-TNBs + US in inhibiting proliferation and inducing apoptosis of tumor cells. Flow cytometry and Western blotting are used to analyse the ROS generation and molecular mechanisms. ICG-TNBs had a particle size of 497.0 ± 29.2 nm and a Zeta potential of -8.05 ± 0.73 mV. In vitro multimodal imaging showed that the image signal intensity of ICG-TNBs increased with concentration. Targeted binding assay confirmed that ICG-TNBs could specifically bind to MCF-7 cells (CXCR4 positive), but not to MDA-MB-468 cells (CXCR4 negative). CCK-8 assay and flow cytometry analysis showed that ICG-TNBs + US could significantly inhibit the growth of MCF-7 breast cancer cells and promote their apoptosis. Flow cytometry and Western blotting showed that ICG-TNBs + US could significantly raise generation of ROS, reduce the expression of CXCR4, inhibit phosphorylation of Akt, and increase the expression of Caspase3 and Cleaved-caspase3. This indicated that ICG-TNBs could effectively inhibit and block the SDF-1/CXCR4 pathway, thus leading to the apoptosis of MCF-7 cells. ICG-TNBs can specifically bind to CXCR4 positive breast cancer cells, furthermore inhibit growth and promote apoptosis of breast cancer cells combined with ultrasonic irradiation by blocking the SDF-1/CXCR4 pathway. This study introduces a novel concept, method and mechanism for integration of targeted diagnosis and treatment of breast cancer.
构建携带小分子 CXCR4 拮抗剂 AMD070 和光吸收材料吲哚菁绿（ICG）的靶向纳米气泡，并研究其体外多模态成像以及抑制乳腺癌细胞生长的机制和效果。通过碳二亚胺反应和机械振荡构建携带 AMD070 和 ICG 的纳米气泡（ICG-TNBs）。测定其物理特性和体外多模态成像。激光共聚焦显微镜观察 ICG-TNBs 与人乳腺癌细胞的结合潜力。CCK-8 和流式细胞术分析 ICG-TNBs+US 抑制肿瘤细胞增殖和诱导细胞凋亡的作用。流式细胞术和 Western blot 分析 ROS 生成和分子机制。ICG-TNBs 的粒径为 497.0±29.2nm，Zeta 电位为-8.05±0.73mV。体外多模态成像显示，ICG-TNBs 的图像信号强度随浓度增加而增加。靶向结合实验证实，ICG-TNBs 可以特异性结合 MCF-7 细胞（CXCR4 阳性），但不能结合 MDA-MB-468 细胞（CXCR4 阴性）。CCK-8 测定和流式细胞术分析表明，ICG-TNBs+US 可显著抑制 MCF-7 乳腺癌细胞的生长并促进其凋亡。流式细胞术和 Western blot 显示，ICG-TNBs+US 可显著提高 ROS 的产生，降低 CXCR4 的表达，抑制 Akt 的磷酸化，并增加 Caspase3 和 Cleaved-caspase3 的表达。这表明 ICG-TNBs 可以有效抑制和阻断 SDF-1/CXCR4 通路，从而导致 MCF-7 细胞凋亡。ICG-TNBs 可以特异性结合 CXCR4 阳性乳腺癌细胞，通过阻断 SDF-1/CXCR4 通路，结合超声辐射进一步抑制和阻断乳腺癌细胞的生长并促进其凋亡。本研究为乳腺癌的靶向诊断和治疗一体化引入了新的概念、方法和机制。
