CAJ | 학술논문

目的探讨甲胎蛋白(AFP)靶向超小型超顺磁性氧化铁(AFP-USPIO)分子探针在肝细胞癌(HCC)主动靶向特异性成像方面的价值.方法表面修饰的USPIO连接抗AFP抗体后构建AFP-USPIO靶向分子探针.实验分为AFP-USP1O组(靶向组)和USPIO组(非靶向组).将铁浓度为100 μg/ml的AFP-USPIO及USPIO,按照150μl、100 μl、50 μl的剂量分别与HepG2细胞孵育4h 后行体外磁共振成像,并测量T2WI信噪比(SNR).构建原位肝癌动物模型并随机分为两组(每组5只),分别于尾静脉注射AFP-USPIO或USPIO,注射剂量20μmol/kg.注射前及注射后1h进行磁共振成像,测量T2WI肝脏肿瘤与周围肝组织的对比噪声比(CNR),计算增强前后及两组之间CNR 的差异.普鲁士蓝铁染色验证SNR、CNR变化与细胞及组织铁含量的关系.结果细胞学实验显示两组体外磁共振SNR均随AFP-USPIO或USPIO剂量的增加而下降,3种剂量间(P＜0.05)以及同一剂量时两组间SNR差异均具有统计学意义(P＜0.05).普鲁士蓝染色结果显示靶向组细胞内铁颗粒随剂量的增加而增多,并与SNR呈负相关关系(P=0.00,r=-0.926),而非靶向组铁染色的细胞数则明显降低.在注射AFP-USP1O前后,大鼠肝脏肿瘤的CNR分别为2.05±0.88和0.96±0.31,差异有统计学意义(P=0.028,t=3.380);而注射USPIO前后肝脏肿瘤的CNR分别为2.25±1.50和2.57±1.49,差异无统计学意义(P=0.275,t=1.263).靶向与非靶向组,肿瘤强化后的CNR差异也具有统计学意义(P=0.042,t=3.487).病理结果显示靶向组肿瘤组织内见大量铁染色颗粒,而非靶向组铁染色颗粒则明显减少.结论 AFP-USPIO分子探针对HepG2细胞及大鼠模型原位肝癌具有良好的主动靶向作用,可实现肝癌的特异性成像和诊断.
목적 탐토갑태단백(AFP)파향초소형초순자성양화철(AFP-USPIO)분자탐침재간세포암(HCC)주동파향특이성성상방면적개치.방법 표면수식적USPIO련접항AFP항체후구건AFP-USPIO파향분자탐침.실험분위AFP-USP1O조(파향조)화USPIO조(비파향조).장철농도위100 μg/ml적AFP-USPIO급USPIO,안조150μl、100 μl、50 μl적제량분별여HepG2세포부육4h 후행체외자공진성상,병측량T2WI신조비(SNR).구건원위간암동물모형병수궤분위량조(매조5지),분별우미정맥주사AFP-USPIO혹USPIO,주사제량20μmol/kg.주사전급주사후1h진행자공진성상,측량T2WI간장종류여주위간조직적대비조성비(CNR),계산증강전후급량조지간CNR 적차이.보로사람철염색험증SNR、CNR변화여세포급조직철함량적관계.결과 세포학실험현시량조체외자공진SNR균수AFP-USPIO혹USPIO제량적증가이하강,3충제량간(P＜0.05)이급동일제량시량조간SNR차이균구유통계학의의(P＜0.05).보로사람염색결과현시파향조세포내철과립수제량적증가이증다,병여SNR정부상관관계(P=0.00,r=-0.926),이비파향조철염색적세포수칙명현강저.재주사AFP-USP1O전후,대서간장종류적CNR분별위2.05±0.88화0.96±0.31,차이유통계학의의(P=0.028,t=3.380);이주사USPIO전후간장종류적CNR분별위2.25±1.50화2.57±1.49,차이무통계학의의(P=0.275,t=1.263).파향여비파향조,종류강화후적CNR차이야구유통계학의의(P=0.042,t=3.487).병리결과현시파향조종류조직내견대량철염색과립,이비파향조철염색과립칙명현감소.결론 AFP-USPIO분자탐침대HepG2세포급대서모형원위간암구유량호적주동파향작용,가실현간암적특이성성상화진단.
Objective To investigate the potential of AFP-targeted ultrasmall superparamagnetic particles of iron oxide (USPIO) molecular probe in specific detection of hepatocellular carcinoma (HCC) with MRI.Methods The targeted probe was synthesized by conjugating AFP antibody with modified USPIO.Two groups treated with AFP-USPIO and USPIO were set up in the study.The HepG2 cells were incubated with AFP-USPIO or USPIO (100 μg/ml) respectively with the dosage of 50 μ1,100 μl or 150 μl for 4 hours,followed by MR imaging in vitro.The signal-noise ratio (SNR) of the cells on T2-weighted image (T2WI) was measured.The rat models with orthotopic HCC were divided into two groups with 5 rats for each group at random.Pre-and post-contrast enhanced (after 1 hour) MR imaging were performed with caudal vein injection at a dosage of 20μg/ml.The contrast noise ratio (CNR) on T2WI and the difference of CNR between pre-and post-enhancement or between both groups were calculated.The relationship of SNR or CNR with the iron particles in cells or tumors was confirmed by Prussian blue iron staining.Results Cytology experiment showed the SNR in both groups was decreased with the increase of the dosage of AFP-USPIO or USPIO,indicating statistically significantdifference in SNR among three different doseage groups (P＜0.05).Prussian blue iron staining showed that the iron particles in cells were increased with the increase of AFP-USPIO dosage,and was negatively correlated with SNR (P=0.00,r=-0.926).However,the iron particles were less in cells in USPIO group.The CNRs of liver tumors in Wistar rat of pre-and post-AFP-USPIO injection were 2.05±0.88 and 0.96±0.31 respectively,indicating a significant difference (P=0.028,t=3.380).However,the CNRs in USPIO group,2.25±1.50 and 2.57±1.49,showed no statistical difference (P=0.275,t=1.263).The CNR after enhancement also had a statistical difference between both groups(P=0.042,t=3.487).Pathological results confirmed more iron particles in tumor tissues in AFP-USPIO group,whereas less in USPIO group.Conclusion AFP-USPIO molecular probes can initiatively target to the HepG2 cells and the liver cancer of rat models expressing AFP,which may help to achieve the specificity of MR imaging in the diagnosis of HCC.