Development of a Pancreatic Cancer Specific Binding Peptide Using Phage Display
- Author:
Dong Won LEE
1
;
Jae Myung PARK
;
Seung Mok YANG
;
Moon Hwa KWAK
;
Yoon Jin ROH
;
In Seok LEE
;
Myung Gyu CHOI
Author Information
- Publication Type:Original Article
- Keywords: Bacteriophages; Pancreatic neoplasms; Peptides; Photochemotherapy
- MeSH: Bacteriophages; Cell Line; Cell Survival; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Fluorescein; Fluorescence; Fluorescent Antibody Technique; Humans; Immunohistochemistry; Models, Animal; Pancreatic Neoplasms; Peptides; Photochemotherapy; Prognosis; Survival Rate
- From:The Korean Journal of Gastroenterology 2019;74(1):30-41
- CountryRepublic of Korea
- Language:Korean
- Abstract: BACKGROUND/AIMS: Pancreatic cancer has a very poor prognosis, and early diagnosis is a way to increase the survival rate of patients. The purpose of this study was to develop pancreatic cancer-specific peptides for imaging studies. METHODS: Three pancreatic cancer cell lines, MIA PaCa-2, UACC-462, and BxPC-3, and a control cell line, CCD841, were used. Biopannings were performed on MIA PaCa-2 using a phage display library. After this, the peptides were synthesized and labeled with fluorescein isothiocyanate (FITC). Immunocytochemistry (ICC), enzyme-linked immunosorbent assay (ELISA), and fluorescence-activated cell sorter (FACS) were performed to examine the specific binding. To examine its therapeutic applications, a photosensitizer, chlorin e6 (Ce6), was conjugated on the peptide and photodynamic therapy was performed. Cell survival was investigated using a [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assay. RESULTS: After three biopannings, the phages were amplified from 1.4×104 to 3.2×105 plaque-forming units. The most strongly binding phage was selected from the ELISA and ICC results. FITC-labeled peptide, M5, in the three pancreatic cancer cell lines showed significantly higher immunofluorescence in the ICC experiments than that of CCD841. The higher binding ability to MIA PaCa-2 cells was confirmed from FACS analysis, which showed a right shift compared to CCD841. M5 bound to Ce6 showed a significantly lower cell survival rate than that of Ce6 alone in photodynamic therapy, which was observed consistently as a change in the tumor size and fluorescence intensity in MIA PaCa-2 cell-implanted animal models. CONCLUSIONS: This study showed that the noble peptide, M5, binds specifically to the pancreatic cancer cell line, MIA PaCa-2. The M5 peptide has potential use in future optical diagnostic and therapeutic purposes.
