2024-12-18 作者：ws 来源：文献：
Efficacy evaluation and mechanism study on inhibition of breast cancer cell growth by multimodal targeted fluorescent nanobubbles carrying AMD070 and ICG
文献链接：
https://pubmed.ncbi.nlm.nih.gov/32155591/作者：
Daijia Shen, Lianhua Zhu, Yu Liu1 Yanli Peng, Minmin Lan, Kejing Fang, Yanli Guo
相关产品：
DSPE-PEG2000-COOH（磷脂-聚乙二醇2000-羧基）
原文摘要：
Objective: 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. Methods: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. Results: ICG-TNBs had a particle size of 497.0±29.2 nm and a Zeta potential of -8.05±0.73mV. 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. Conclusion: 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.
DSPE - PEG2000 - COOH 由三部分组成。DSPE（1,2 - 二硬脂酰 - sn - 甘油 - 3 - 磷酸乙醇胺）是一种磷脂，其具有两条长的脂肪酸链，这使得分子的 DSPE 部分具有疏水性。PEG2000（分子量为 2000 的聚乙二醇）是亲水性的聚合物，它作为连接桥将 DSPE 和 COOH（羧基）连接起来。PEG 的存在可以增加分子的水溶性和生物相容性。COOH 代表羧基，是一种具有化学反应活性的官能团，它使得该化合物能够参与多种化学合成反应。该文献基于此，携带AMD070和ICG的靶向荧光纳米气泡（ICG-TNBs）的合成如下：
图：DSPE结构式
DSPE-PEG2000-AMD070的制备：
将DSPE-PEG2000-COOH、NHS和EDC以一定的摩尔比溶解在二甲亚砜中，室温下摇晃激活；活化脂质DSPE-PEG2000-COO-琥珀酰亚胺和AMD070进行酰胺反应。反应结束后，将溶液转移到透析袋中，去除未反应的NHS、EDC·HCl和AMD070，用冷冻干燥器去除DMSO和去离子水，获得固体DSPE-PEG2000-AMD070。将DSPE-PEG2000-COOH、AMD070和DSPE-PEG2000-AMD070溶解于氘化DMSO中，用核磁共振（NMR）谱仪检测。
携带AMD070和ICG的靶向荧光纳米气泡（ICG-TNBs）的构建：
将DPPA、DPPC、DPPE、DPPG、DSPE-PEG2000-AMD070和ICG混合，充分溶解于甘油PBS溶液中，避光保存，卧式摇床孵育过夜，使其完全溶解。将溶液转移到青霉素瓶中，用C3F8填充，以取代瓶中的空气。使用ST银水银胶囊振荡器振荡溶液。振荡后，离心，去除未溶解的脂质和微泡。将获得的icg - tnb放置在冰箱中，以便在合适的时间使用。以DPPA、DPPC、DPPE、DPPG和DSPE-PEG2000-COOH为原料，按照上述方法制备了空白纳米气泡。
图：双荧光法验证图像
结论：
该文献成功制备了基于DSPE-PEG2000-COOH合成的携带AMD070和ICG的靶向荧光纳米气泡（ICG-TNBs）。实验结果证明，ICG-TNBs的图像信号强度随浓度的增加而增加，可以特异性地与MCF-7细胞（CXCR4阳性）结合，说明ICG-TNBs可特异性结合CXCR4 cancer细胞，通过阻断SDF-1/CXCR4通路，联合超声照射抑制cancer细胞生长，促进细胞Apoptosis 。
