파지 디스플레이를 이용한 췌장암 특이적 결합 펩타이드 개발
Development of a Pancreatic Cancer Specific Binding Peptide Using Phage Display
이동원, 박재명, 양승목, 곽문화, 노윤진, 이인석, 최명규,
소속 상세정보
이동원 ( Lee Dong-Won )
가톨릭광의학연구소
박재명 ( Park Jae-Myung )
가톨릭대학교 의과대학 서울성모병원 내과
양승목 ( Yang Seung-Mok )
가톨릭광의학연구소
곽문화 ( Kwak Moon-Hwa )
가톨릭광의학연구소
노윤진 ( Roh Yoon-Jin )
가톨릭광의학연구소
이인석 ( Lee In-Seok )
가톨릭대학교 의과대학 서울성모병원 내과
최명규 ( Choi Myoung-Gyu )
가톨릭대학교 의과대학 서울성모병원 내과
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.
키워드
Bacteriophages; Pancreatic neoplasms; Peptides; Photochemotherapy
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