Jones Kelvin M, Karanam Balasubramanyam, Jones-Triche Jacqueline, Sandey Maninder, Henderson Henry J, Samant Rajeev S, Temesgen Samuel, Yates Clayton, Bedi Deepa
Department of Biomedical Sciences, College of Veterinary Medicine, Tuskegee University, Tuskegee, AL, United States.
Department of Biology, Center for Cancer Research, Tuskegee University, Tuskegee, AL, United States.
Front Oncol. 2018 Dec 17;8:625. doi: 10.3389/fonc.2018.00625. eCollection 2018.
Epithelial to mesenchymal transition (EMT) is believed to be crucial for primary tumors to escape their original residence and invade and metastasize. To properly define EMT, there is a need for ligands that can identify this phenomenon in tumor tissue and . A phage-display selection screening was performed to select novel binding phage peptides for identification of EMT in breast cancer. Epithelial breast cancer cell line, MCF-7 was transformed to mesenchymal phenotype by TGF-β treatment and was used for selection. Breast fibroblasts were used for subtractive depletion and breast cancer metastatic cell lines MDA-MB-231, T47D-shNMI were used for specificity assay. The binding peptides were identified, and their binding capacities were confirmed by phage capture assay, phage-based ELISA, immunofluorescence microscopy. The phage peptide bearing the 7-amino acid sequence, LGLRGSL, demonstrated selective binding to EMT phenotypic cells (MCF-7/TGF-β and MDA-MB-231) as compared to epithelial subtype, MCF-7, T47D and breast fibroblasts (Hs578T). The selected phage was also able to identify metastatic breast cancer tumor in breast cancer tissue microarray (TMA). These studies suggest that the selected phage peptide LGLRGSL identified by phage-display library, showed significant ability to bind to mesenchymal-like breast cancer cells/ tissues and can serve as a novel probe/ligand for metastatic breast cancer diagnostic and imaging.
上皮-间质转化（EMT）被认为对原发性肿瘤逃离其原发部位并发生侵袭和转移至关重要。为了准确界定EMT，需要能够在肿瘤组织中识别这一现象的配体。进行了噬菌体展示筛选以选择用于识别乳腺癌中EMT的新型结合噬菌体肽。通过转化生长因子-β（TGF-β）处理将上皮性乳腺癌细胞系MCF-7转化为间充质表型，并用于筛选。乳腺成纤维细胞用于消减性去除，乳腺癌转移细胞系MDA-MB-231、T47D-shNMI用于特异性测定。鉴定了结合肽，并通过噬菌体捕获测定、基于噬菌体的酶联免疫吸附测定、免疫荧光显微镜检查确认了它们的结合能力。与上皮亚型MCF-7、T47D和乳腺成纤维细胞（Hs578T）相比，带有七氨基酸序列LGLRGSL的噬菌体肽表现出对EMT表型细胞（MCF-7/TGF-β和MDA-MB-231）的选择性结合。所选噬菌体还能够在乳腺癌组织微阵列（TMA）中识别转移性乳腺癌肿瘤。这些研究表明，通过噬菌体展示文库鉴定的所选噬菌体肽LGLRGSL显示出与间充质样乳腺癌细胞/组织结合的显著能力，并且可以作为转移性乳腺癌诊断和成像的新型探针/配体。
