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| Status |
Public on Dec 30, 2015 |
| Title |
Generation of a monoclonal antibody recognizing the CEACAM glycan structure and inhibiting adhesion using cancer tissue-originated spheroid as an antigen |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Spheroids cultured directly from tumours can better reflect in vivo tumour characteristics than two-dimensional monolayer culture or three-dimensional culture of established cell lines. In this study, we generated antibodies by directly immunizing mice with cancer tissue-originated spheroids from colorectal cancer. We performed phenotypic screening via recognition of the surface of the spheroids and inhibition of their adhesion to extracellular matrices to identify a monoclonal antibody, clone 5G2. The antibody inhibited cell migration in two-dimensional culture and promoted cell detachment. Western blotting and immunohistochemistry detected the 5G2 signal in many colorectal cancer spheroids, as well as patient tumours, but the signal was reduced in various cell lines. The expression pattern also changed when spheroids were cultured in two-dimensional conditions. We found that 5G2 recognized the N-glycan structure, including Lea and Lec, and their major carrier proteins were CEACAM5 and CEACAM6. Translocation of integrin 4 from the lateral membrane to the extracellular matrix contact interface in the spheroids was delayed by pre-incubation with 5G2. Thus, cancer tissue-originated spheroids can be a useful antigen for generating novel anti-cancer antibodies as the glycan structure is lost in cell lines but retained in cancer tissue-originated spheroids.
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| Overall design |
Gene expression was compared in cancer tissue-originated spheroids (CTOSs) under floating conditions, under gel embeded conditions, CTOS-derived xenograft tumors, and corresponding microdissected patient tumors. One experiment was performed for each sample.
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| Contributor(s) |
Inoue M |
| Citation(s) |
27098764 |
| Submission date |
Dec 29, 2015 |
| Last update date |
Nov 27, 2018 |
| Contact name |
Masahiro Inoue |
| E-mail(s) |
masa_inoue@kuhp.kyoto-u.ac.jp
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| Organization name |
Kyoto Univ Graduate School of Medicine
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| Department |
Clinical Bio-resource Research and Development
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| Street address |
Med-Pharm Collaboration Bldg 503, Shimoadachi-cho 46, Sakyou-ku, Kyoto, 606-8304, Japan
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| City |
Kyoto |
| State/province |
Kyoto |
| ZIP/Postal code |
606-8304 |
| Country |
Japan |
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| Platforms (1) |
| GPL13607 |
Agilent-028004 SurePrint G3 Human GE 8x60K Microarray (Feature Number version) |
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| Samples (32)
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| Relations |
| BioProject |
PRJNA307193 |
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