标题：TLR5 is a new reporter for triple-negative breast cancer indicated by radioimmunoimaging and fluorescent staining
作者：Shi, Dai; Liu, Weiwei; Zhao, Shanshan; Zhang, Chao; Liang, Ting; Hou, Guihua
作者机构：[Hou, Guihua] Shandong Univ, Key Lab Expt Teratol, Minist Educ, Jinan, Shandong, Peoples R China.; [Hou, Guihua] Shandong Univ, Biomed Isotope Res C 更多
通讯作者：Hou, GH;Hou, GH
通讯作者地址：[Hou, GH]Shandong Univ, Key Lab Expt Teratol, Minist Educ, Jinan, Shandong, Peoples R China;[Hou, GH]Shandong Univ, Biomed Isotope Res Ctr, Sch Med, J 更多
来源：JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
关键词：fluorescence imaging; Phosphorautoradiography; Radioiodine 125; TLR5;; triple-negative breast cancer
摘要：Triple-negative breast cancer (TNBC) is a highly aggressive tumour that lacks marker for targeted diagnosis. Recently, it was reported that toll-like receptor 5 (TLR5) was associated with some kind of tumours, especially in TNBC, but whether it could be used as a non-invasive monitoring target is not fully understood. Here, we established TLR5(-) 4T1 cell line with lentivirus-shRNA-TLR5 knock-down transfection (with tag GFP, green fluorescent protein, TLR5(-) 4T1) and control TLR5(+) 4T1 cell line with negative control lentivirus transfection. The effect of TLR5 down-regulation was detected with qPCR and Western blot. I-125-anti-TLR5 mAb and control isotype I-125-IgG were prepared and injected to TLR5(+/-) 4T1-bearing mice models, respectively. Whole-body phosphor-autoradiography, fluorescence imaging and biodistribution were performed. Furthermore, ex vivo tumour TLR5 expression was proved through immunohistochemistry staining. We found that I-125-anti-TLR5 mAb could bind to TLR5(+) 4T1 with high affinity and specificity. Whole-body phosphor-autoradiography after I-125-anti-TLR5 mAb injection showed TLR5(+) 4T1 tumour images in 24 hours, more clearly in 48 hours. Radioactivities in tumour tissues were positively related with TLR5 expression. Biodistribution assay showed that I-125-anti-TLR5 mAb was mainly metabolized through the liver and kidney, and I-125-anti-TLR5 mAb was much more accumulated in TLR5(+) 4T1 tumour than TLR5(-) 4T1. In vivo fluorescence imaging successfully showed tumour tissues clearly both in TLR5(+) and TLR5(-) 4T1 mice compared with lentivirus untreated 4T1 tumour. Immunohistochemistry staining showed that TLR5 expression in tumours was indeed down-regulated in TLR5(-) 4T1 mice. Our results indicated that I-125-antiTLR5 mAb was an ideal agent for non-invasive imaging of TLR5(+) tumours; TLR5 may be as a novel molecular target for TNBC non-invasive diagnosis.