靶向乳腺癌细胞的多功能纳米分子探针介导光声成像及治疗

Targeted multifunctional nanoparticles probe mediated photoacoustic imaging and treatment study of breast carcinoma in vitro

目的制备一种载Fe3O4同时靶向乳腺癌的多功能纳米分子探针，观察其体外光声成像效果，并探讨其体外寻靶能力及光声介导乳腺癌细胞凋亡的能力。方法采用双乳化法和碳二亚胺法，制备载Fe3O4同时连接靶向乳腺癌单克隆抗体（Herceptin）的高分子多功能纳米分子探针，检测其一般物理特性、Herceptin与纳米粒的连接情况及其体外寻靶能力；对不同浓度的纳米粒进行光声成像；体外评价靶向纳米粒介导光声对乳腺癌细胞的抑制作用。结果成功制备的靶向载Fe3O4多功能纳米分子探针平均粒径为（235．40±53．75）nm，Zeta电位（-13．4±4．7）mv；Fe3O4颗粒分布于纳米探针壳层；间接免疫荧光法观察到抗体成功连接到纳米探针表面；体外寻靶能力实验显示乳腺癌SKBR3细胞株周围有大量靶向纳米探针环绕。体外光声成像显示随Fe3O4含量增加，其光声信号逐渐增强。体外细胞抑制实验显示激光辐照后的乳腺癌细胞凋亡率明显升高，证明制备的纳米探针能够协同光声对乳腺癌细胞产生明显的抑制作用。结论制备的靶向载Fe3O4多功能纳米分子探针可作为光声成像造影剂，同时在光声介导下，能够显著抑制乳腺癌细胞的增殖。

Objective To investigate the ability of targeting in vitro breast carcinoma and photoacoustic-mediate apoptosis of breast carcinoma of the nanoparticle probe loaded with Fe3O4 and observe its consequence of photoacoustic imaging in vitro. Methods The polymeric multifunctional nanoparticles probe that was loaded with Fe3O4 and connected with Herceptin targeting breast carcinoma was prepared in the use of double emulsion method and carbodiimide method. General physical property, the condition of Herceptin connected with nanoparticles and the ability of targeting of the probe were tested. The different concentration of nanoparticles was imaged by photoacoustic. The inhibiting effect of targeting nanoparticles on breast carcinoma cells was evaluated in vitro. Results The targeting polymeric multifunctional nanoparticles probes loaded with Fe3O4 were prepared successfully with average particle diameter of （235.4±53.75）nm and Zeta potential （-13.4±4.7）inV. Fe3 04 particles dispersed on the shell of the probes. Antibody Herceptin was successfully connected with the surface of the nanoprobes. There were massive targeting nanoprobes surround the breast carcinoma cell strains SKBR3 in the targeting group in vitro. The photoascoustic signal of the nanoprobes enhanced with the increase of Fe3O4 concentration in photoacoustic experiment in vitro. The apoptotic rate of breast carcinoma increased after the laser irradiation in the cell inhibition experiment in vitro. That proved the obvious inhibition of breast carcinoma cells was caused by photothermal effect of the prepared nanoprobes. Conclusions The prepared polymeric multifunctional nanoparticles probe that was loaded with Fe3O4 and connected with Herceptin targeting breast carcinoma can be used as a photoacoustic contrast agent, which can inhibit the proliferation of breast carcinoma by targeting photoacoustic therapy.