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Archive > Volume 20 Issue 5 > 2013,20(5):515-521. DOI:10.3872/j.issn.1007-385X.2013.05.002 Prev Next
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Screening and identifying of homing peptides to bladder cancer BIU-87 cells in Chinese
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Abstract:
Objective: To screen a peptide strongly binding to the bladder cancer BIU-87 cells from phage 7-mer cyclic peptide library in vitro in Chinese, and identify its specificity. Methods: 3 rounds subtraction screening was performed on a phage 7-mer cyclic peptide library, with the BIU-87 cells as the target cells and normal human bladder epithelial cells as the absorber cells. Phage clones which can positively bind to BIU-87 cells were identified by ELISA, and those coding DNA were sequenced to analyze the homology. Strongly positive peptide was chemically synthetized to prepare fluorescent probe by taging fluorescein isothiocyanate (FITC). The affinity between the fluorescent probes and BIU- 87 bladder cancer cells, human normal bladder epithelial cells, human prostate carcinoma PC3M cells, human hepatocellular carcinoma SMMC-7721 cells and human colon cancer HCT116 cells were identified by flow cytometry and fluorescence microscopy.Results:The c7c phage-display peptides library was enriched for 25 times through 3 rounds of subtraction screening and the positive rate was 76%. 10 strongly positive clones were obtained. For the 10 strongly positive clones, the amino acid sequences of SISSLTH, MARYMSA and TVRTSAD were consensus sequence. The binding rate of small molecular fluorescent probe FITC-SISSLATH binding to the BIU- 87 bladder cancer cells (80.06±8.78)% was obviouly higher than of FITC-MARYMSA(52.93±7.28)%, FITC-TVRTSAD(38.04±7.47)%, FITC -EDRKETA(1.91±1.37)% and FITC(9.85±2.9)% (all P<0.01). The binding rate of the BIU- 87 bladder cancer cells binding to FITC-SISSLATH(80.06±8.78)% was higher than that of the normal human bladder epithelial cells (13.89±197)%, human prostate carcinoma PC3M cells(8.13±2.85)%, human hepatocellular carcinoma SMMC-7721 cells (27±2.87)% and human colon cancer HCT116 cells(2.33±1.75)%(all P<0.01). Conclusion: A targeting peptide SISSLTH binding to BIU-87 cells with high specificity and efficiency was obtained from the phage 7-mer cyclic peptide library through 3 rounds of subtraction screening in vitro.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81172744)
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