Department of Urology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510700, Guangdong, PR China.
J Exp Clin Cancer Res. 2011 Nov 10;30(1):105. doi: 10.1186/1756-9966-30-105.
Specific peptide ligands to cell surface receptors have been extensively used in tumor research and clinical applications. Phage display technology is a powerful tool for the isolation of cell-specific peptide ligands. To screen and identify novel markers for renal cell carcinoma, we evaluated a peptide that had been identified by phage display technology.
A renal carcinoma cell line A498 and a normal renal cell line HK-2 were used to carry out subtractive screening in vitro with a phage display peptide library. After three rounds of panning, there was an obvious enrichment for the phages specifically binding to the A498 cells, and the output/input ratio of phages increased about 100 fold. A group of peptides capable of binding specifically to the renal carcinoma cells were obtained, and the affinity of these peptides to the targeting cells and tissues was studied.
Through a cell-based ELISA, immunocytochemical staining, immunohistochemical staining, and immunofluorescence, the Phage ZT-2 and synthetic peptide ZT-2 were shown to specifically bind to the tumor cell surfaces of A498 and incision specimens, but not to normal renal tissue samples.
A peptide ZT-2, which binds specifically to the renal carcinoma cell line A498 was selected from phage display peptide libraries. Therefore, it provides a potential tool for early diagnosis of renal carcinoma or targeted drug delivery in chemotherapy.
细胞表面受体的特定肽配体已被广泛用于肿瘤研究和临床应用。噬菌体展示技术是分离细胞特异性肽配体的有力工具。为了筛选和鉴定肾细胞癌的新型标志物,我们评估了一种通过噬菌体展示技术鉴定的肽。
用噬菌体展示肽库对肾癌细胞系 A498 和正常肾细胞系 HK-2 进行体外消减筛选。经过三轮淘选,能够特异性结合 A498 细胞的噬菌体明显富集,噬菌体的输出/输入比值增加了约 100 倍。获得了一组能够特异性结合肾癌细胞的肽,并研究了这些肽与靶细胞和组织的亲和力。
通过细胞 ELISA、免疫细胞化学染色、免疫组织化学染色和免疫荧光染色,噬菌体 ZT-2 和合成肽 ZT-2 被证明特异性结合 A498 肿瘤细胞表面和切口标本,但不结合正常肾组织标本。
从噬菌体展示肽库中筛选出一种特异性结合肾癌细胞系 A498 的肽 ZT-2。因此,它为肾细胞癌的早期诊断或化疗中的靶向药物输送提供了一种潜在的工具。