Fluorescent imaging of superficial head and neck squamous cell carcinoma using a γ-glutamyltranspeptidase-activated targeting agent: a pilot study.
Fluorescent imaging of superficial head and neck squamous cell carcinoma using a γ-glutamyltranspeptidase-activated targeting agent: a pilot study.
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DOI：
10.1186/s12885-016-2421-z
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发表时间：
2016-07-07
期刊：
影响因子：
3.8
通讯作者：
Urano Y
中科院分区：
文献类型：
--
作者：
Mizushima T;Ohnishi S;Shimizu Y;Hatanaka Y;Hatanaka KC;Hosono H;Kubota Y;Natsuizaka M;Kamiya M;Ono S;Homma A;Kato M;Sakamoto N;Urano Y
头颈部浅表鳞状细胞癌(HNSCC)的内窥镜检查具有挑战性，因为其形态特征有限，而且由于严重的粘膜刺激，碘不能应用于头颈部病变。γ-谷氨酰转肽酶(GGT)是一种细胞表面酶，在多种肿瘤中过表达，有报道称γ-谷氨酰羟甲基罗丹明绿(GGLHMRG)是一种可被酶激活的荧光靶向剂，在GGT特异性序列被切割后几分钟内即可被激活。我们研究了是否可以通过将gGlu-HMRG应用于临床样本来检测早期HNSCC。GGlu-HMRG作用于4株HNSCC细胞，用荧光显微镜和流式细胞仪观察荧光。采用免疫组织化学方法对近期3例内窥镜下黏膜下剥离(ESD)中GGT的表达进行了研究。对8例经ESD或手术切除的临床标本进行gGlu-HMRG荧光成像，前瞻性测量肿瘤和正常黏膜感兴趣区的荧光强度。所有四个应用gGlu-HMRG的细胞系都发出绿色荧光。免疫组织化学检测显示，GGT在最近3例ESD患者的HNSCC中高表达，而在正常粘膜中几乎不表达。荧光成像显示，在所有8个切除的肿瘤中，应用gGlu-HMRG后几分钟内，碘排尿病变就变成了荧光。应用gGlu-HMRG后5分钟，肿瘤ROI荧光强度明显高于正常黏膜。GGlu-HMRG荧光成像有助于HNSCC的早期发现。
Detecting superficial head and neck squamous cell carcinoma (HNSCC) by endoscopy is challenging because of limited morphological hallmarks, and iodine cannot be applied to head and neck lesions due to severe mucosal irritation. γ-glutamyltranspeptidase (GGT), a cell surface enzyme, is overexpressed in several cancers, and it has been reported that γ-glutamyl hydroxymethyl rhodamine green (gGlu-HMRG), a fluorescent targeting agent which can be enzymatically activated and becomes fluorescent after cleavage of a GGT-specific sequence, can be activated within a few minutes after application to animal models. We investigated whether early HNSCC can be detected by applying gGlu-HMRG to clinical samples. gGlu-HMRG was applied to four HNSCC cell lines, and fluorescence was observed by fluorescence microscopy and flow cytometry. Immunohistological examination was performed in three recent cases of endoscopic submucosal dissection (ESD) to investigate GGT expression. Fluorescence imaging with gGlu-HMRG in eight clinical samples resected by ESD or surgery was performed, and fluorescence intensity of tumor and normal mucosa regions of interest (ROI) was prospectively measured. All four gGlu-HMRG-applied cell lines emitted green fluorescence. Immunohistological examination demonstrated that GGT was highly expressed in HNSCC of the recent three ESD cases but barely in the normal mucosa. Fluorescence imaging showed that iodine-voiding lesions became fluorescent within a few minutes after application of gGlu-HMRG in all eight resected tumors. Tumor ROI fluorescence intensity was significantly higher than in the normal mucosa five minutes after gGlu-HMRG application. Fluorescence imaging with gGlu-HMRG would be useful for early detection of HNSCC.