【摘要】 目的：制备针对人表皮受体-2(HER-2)过表达的甲状腺癌靶向性磁共振分子成像纳米探针钆特酸葡胺-帕妥珠单抗(Pertuzumab)(Gd-DOTA-pertuzumab)，通过检测纳米探针Gd-DOTA-pertuzumab的表征、荷瘤裸鼠磁共振成像(MRI)，探究其应用于甲状腺癌MRI的可能性。方法：将Pertuzumab与钆剂偶联，制备HER-2过表达的甲状腺癌靶向性磁共振分子探针Gd-DOTA-pertuzumab。取1,4,7,10-四氮杂环十二烷-1,4,7,10-四乙酸(DOTA)溶液，加入1,2-二氯乙烷(EDC)和N-羟基丁二酰亚胺(NHS)活化后，加入Pertuzumab，混合后在室温下振荡反应过夜，透析得到DOTApertuzumab。加入六水合氯化钆(GdCl3-6H2O)，室温下孵育6 h后加入牛血清白蛋白(BSA)，经超声振荡5 min后得到BSA包裹的纳米探针Gd-DOTA-pertuzumab。选用3～4周龄雌性balb/c裸鼠，在裸鼠背部注射甲状腺癌细胞进行荷瘤，观察该纳米探针在荷瘤裸鼠体内肿块信号。结果：纳米探针Gd-DOTA-pertuzumab的平均水动力尺寸为(117.52±6.23)nm，平均电位为(-23.46±3.57)m V，经透射电子显微镜下显示纳米探针Gd-DOTA-pertuzumab平均颗粒大小为(91.42±13.36)nm，弛豫率为24.18 mmol-1s-1。激光共聚焦显示纳米探针Gd-DOTA-pertuzumab与细胞表面结合。磁共振体外成像显示随着纳米探针Gd-DOTA-pertuzumab的浓度增加，磁共振T1WI信号逐渐升高。清晰的观察到该探针在荷瘤裸鼠体内肿块信号增加，并与周围组织边界清楚。结论：成功合成的纳米探针Gd-DOTA-pertuzumab具有良好的MRI特点，可与目的细胞靶向性结合，在甲状腺癌MRI诊断中具有重要价值。更多还原

【Abstract】 Objective: To prepare the targeted molecular probe Gd-DOTA-pertuzumab of magnetic resonance imaging(MRI) for overexpressed human epidermal receptor(HER-2), and to explore the feasibility of that in MRI application of thyroid cancer through detected the representation of nanoprobe gadolinium-DOTA-pertuzumab(GdDOTA-pertuzumab) and the MRI in nude mouse bearing the tumor. Methods: Pertuzumab was coupled with Gd,and then the targeted MRI molecular probe Gd-DOTA-pertuzumab for overexpressed human epidermal receptor(HER-2) was prepared. 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic-acid(DOTA) solution was excited after 1,2-Dichloroethane(EDC) and N-Hydroxysuccinimide(NHS) were added, and pertuzumab was added into them after that. The reaction of whole reaction system overnighted under room temperature after the system was mixed,and then the DOTA-pertuzumab was obtained through dialysis. After gadolinium chloride hexahydrate(GdCl3-6H2O)was added into the reaction system, the new reaction system was incubated within 6h under room temperature, and then the bovine serum albumin(BSA) was added into them. After that, we obtained the encapsulated nanoprobe GdDOTA-pertuzumab with BSA post 5min under sonic oscillation. The female balb/c nude mice at 3-4 weeks of age were selected, and the thyroid cancer cells were injected on the back of nude mice to bear cancer. Finally, the signal of the nanoprobe in the mass within the body of nude mice with bearing cancer were observed. Results: The hydrodynamic size of the nanoprobe Gd-DOTA-pertuzumab was(117.52±6.23) nm, and the electric potential was(-23.46±3.57)mV. The size of nanoprobe Gd-DOTA-pertuzumab that was showed under transmission electron microscopy was(91.42±13.36) nm, and the relaxation rate was 24.18 mmol-1s-1. The results of laser confocal displayed the nanoprobe Gd-DOTA-pertuzumab was bound to cell surface. The results of in vitro MRI showed that T1WI signal value increased gradually with the increasing of the concentration of nanoprobe Gd-DOTA-pertuzumab. It was clear that the signal values of the observed probe increased in the mass within the body of nude mice with bearing cancer, which was clear with the boundary of periphery tissue. Conclusion: The successfully synthesized Gd-DOTA-pertuzumab has favorable MRI characteristics and it can combined with target cells. It has important value in MRI diagnosis of thyroid cancer.更多还原