基于c-Met抗体的近红外荧光探针用于肝癌的活体成像研究  

c-Met antibody-conjugated near-infrared fluorescent probe for in vivo imaging of hepatocellular carcinoma xenograft models

作　　者：易柳彤 杨美林 宫娜娜[4] 余美红 曹婉维[1] 李展宇[1] 李丹 Yi Liutong;Yang Meilin;Gong Nana;Yu Meihong;Cao Wanwei;Li Zhanyu;Li Dan(Department of Pathology,the Fifth Affiliated Hospital of Sun Yat-sen University,Guangdong Zhuhai 519000,China;Guangdong Provincial Key Laboratory of Biomedical Imaging,the Fifth Affiliated Hospital of Sun Yat-sen University,Guangdong Zhuhai 519000,China;Guangdong Provincial Engineering Research Center of Molecular Imaging,the Fifth Affiliated Hospital of Sun Yat-sen University,Guangdong Zhuhai 519000,China;Department of Clinical Laboratory,the Fifth Affiliated Hospital of Sun Yat-sen University,Guangdong Zhuhai 519000,China)

机构地区：[1]中山大学附属第五医院病理科,广东珠海519000 [2]中山大学附属第五医院广东省生物医学影像重点实验室,广东珠海519000 [3]中山大学附属第五医院广东省分子影像技术工程研究中心,广东珠海519000 [4]中山大学附属第五医院检验科,广东珠海519000

出　　处：《中华介入放射学电子杂志》2021年第4期407-414,共8页Chinese Journal of Interventional Radiology：electronic edition

基　　金：中山大学附属第五医院“五个五”工程青年人才项目(2016972024656)。

摘　　要：目的探靶向近红外荧光成像为肝癌的精准诊疗带来了重要的契机,而且细胞间质表皮转化因子(c-Met)在肝癌中的表达显著高于正常肝组织。因此,本研究拟采用靶向c-Met的近红外荧光探针SHRmAb-IR800进行肝癌的活体成像研究。方法从TCGA和HPA数据库中获取并分析c-Met mRNA和蛋白的表达信息。通过Western Blot分析人肝癌细胞系中c-Met蛋白的表达情况。采用流式细胞技术和共聚焦成像实验评估SHRmAb-IR800与肝癌细胞的体外特异结合能力。构建肝癌皮下瘤模型,进行活体成像分析,并量化SHRmAb-IR800的成像肿瘤背景比。结果通过数据库分析发现肝癌中的c-Met mRNA和蛋白表达增加。Western Blot分析也显示人肝癌细胞系表达c-Met蛋白。流式细胞术和共聚焦成像实验均显示探针能特异性靶向c-Met阳性的人源肝癌细胞株;肝癌皮下瘤模型的近红外荧光成像显示靶向探针在皮下瘤的聚集显著高于对照探针(P<0.05),并且探针在96 h的成像肿瘤背景比达到(2.01±0.18)。结论本研究发现靶向c-Met近红外探针能够特异识别c-Met阳性的肝癌细胞和组织,有助于肝癌的诊断,在肝癌的手术导航方面具有潜在应用价值。Objective Targeting near-infrared fluorescent imaging has facilitated the precise diagnosis and treatment of hepatocellular carcinoma(HCC),and cellular mesenchymal-epithelial transition factor(c-Met)has been reported as a highly expressed marker in HCC tissues compared with normal liver tissues.Thus,we intend to use the c-Met targeted near-infrared fluorescent probe SHRmAb-IR800 for imaging in HCC xenograft models.Methods The expression information of c-Met mRNA and protein was obtained and analyzed from TCGA and HPA databases,respectively.The expression of c-Met protein in HCC cell lines was analyzed by Western Blot.Flow cytometry and confocal imaging experiments were used to evaluate the specific binding ability of SHRmAb-IR800 with HCC cells in vitro.HCC subcutaneous xenograft models were constructed to perform in vivo imaging analysis and quantify the imaging tumor background ratio(TBR)of SHRmAb-IR800.Results The data extracted and analyzed from TCGA and HPA showed c-Met mRNA and protein were highly expressed in HCC tissues.Western Blot analysis also displayed that HCC cell lines expressed c-Met protein.Flow cytometry and confocal imaging were performed to verify the high specificity of SHRmAb-IR800 toward human HCC cell lines.Then the in vivo imaging of the probe in HCC xenograft models showed that the aggregation of SHRmAb-IR800 in tumor was higher than the control probe(P<0.05).Moreover,the tumor-to-background ratio(TBR)of the probe in 96 h reached to 2.01±0.18.Conclusions Our study demonstrated that c-Met-targeted probe had highly specific binding ability to HCC cells and tissues,which could be used to detect the HCC and have profound application potential in surgical navigation of HCC.

关 键 词：c-Met(细胞间质表皮转化因子) 近红外荧光探针 肝癌 近红外荧光成像 

分 类 号：R735.7[医药卫生—肿瘤]