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EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy
PURPOSE: The molecular markers cluster of differentiation (CD)24, CD44, adenosine tri-phosphate (ATP) binding cassette protein G2 (ABCG2), and epithelial cell adhesion molecule (EpCAM) are widely used, individually or in combination, to characterize some types of cancer stem cells. In this study we...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Molecular Vision
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3275636/ https://www.ncbi.nlm.nih.gov/pubmed/22328825 |
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author | Mitra, Moutushy Kandalam, Mallikarjuna Harilal, Anju Verma, Rama Shenkar Krishnan, Uma Maheswari Swaminathan, Sethuraman Krishnakumar, Subramanian |
author_facet | Mitra, Moutushy Kandalam, Mallikarjuna Harilal, Anju Verma, Rama Shenkar Krishnan, Uma Maheswari Swaminathan, Sethuraman Krishnakumar, Subramanian |
author_sort | Mitra, Moutushy |
collection | PubMed |
description | PURPOSE: The molecular markers cluster of differentiation (CD)24, CD44, adenosine tri-phosphate (ATP) binding cassette protein G2 (ABCG2), and epithelial cell adhesion molecule (EpCAM) are widely used, individually or in combination, to characterize some types of cancer stem cells. In this study we characterized the EpCAM(+) retinoblastoma (RB) cells for their cancer stem-like properties in vitro. Additionally, we targeted RB tumor cells via redirecting T cells using bispecific EpCAM×CD3 antibody. METHODS: Flow cytometry was used to study the co-expression of EpCAM with putative cancer stem cell markers, such as CD44, CD24, and ABCG2, in RB primary tumors. In vitro methyl thiazol tetrazolium (MTT) assay, invasion assay, and neurosphere formation assay were performed to characterize EpCAM(+) cells for their cancer stem/progenitor cell-like properties. We assessed the in vitro efficacy of bispecific EpCAM×CD3 antibody on RB tumor cell proliferation and validated the results by evaluating effector cytokine production in the culture medium with the ELISA method. RESULTS: EpCAM was co-expressed with all cancer stem cell markers (CD44, CD24, and ABCG2) in primary RB tumors. EpCAM(+) cells showed significantly higher proliferative invasive potential and neurosphere formation in vitro compared to EpCAM(–) Y79 cells. EpCAM(+) cells showed higher β-catenin expression compared to EpCAMˉ cells. EpCAM×CD3 significantly retarded proliferation of RB primary tumor cells. EpCAM×CD3 effectively induced the secretion of effector cytokines, such as interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-10, IL-2, and transforming growth factor (TGF)-β1, and also perforin levels by pre-activated lymphocytes. CONCLUSIONS: EpCAM might be a novel cancer stem cell marker in RB. EpCAM×CD3 antibody redirecting T cells to attack RB tumor cells may prove effective in RB management. Further preclinical studies are needed to confirm the initial findings of our study. |
format | Online Article Text |
id | pubmed-3275636 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Molecular Vision |
record_format | MEDLINE/PubMed |
spelling | pubmed-32756362012-02-10 EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy Mitra, Moutushy Kandalam, Mallikarjuna Harilal, Anju Verma, Rama Shenkar Krishnan, Uma Maheswari Swaminathan, Sethuraman Krishnakumar, Subramanian Mol Vis Research Article PURPOSE: The molecular markers cluster of differentiation (CD)24, CD44, adenosine tri-phosphate (ATP) binding cassette protein G2 (ABCG2), and epithelial cell adhesion molecule (EpCAM) are widely used, individually or in combination, to characterize some types of cancer stem cells. In this study we characterized the EpCAM(+) retinoblastoma (RB) cells for their cancer stem-like properties in vitro. Additionally, we targeted RB tumor cells via redirecting T cells using bispecific EpCAM×CD3 antibody. METHODS: Flow cytometry was used to study the co-expression of EpCAM with putative cancer stem cell markers, such as CD44, CD24, and ABCG2, in RB primary tumors. In vitro methyl thiazol tetrazolium (MTT) assay, invasion assay, and neurosphere formation assay were performed to characterize EpCAM(+) cells for their cancer stem/progenitor cell-like properties. We assessed the in vitro efficacy of bispecific EpCAM×CD3 antibody on RB tumor cell proliferation and validated the results by evaluating effector cytokine production in the culture medium with the ELISA method. RESULTS: EpCAM was co-expressed with all cancer stem cell markers (CD44, CD24, and ABCG2) in primary RB tumors. EpCAM(+) cells showed significantly higher proliferative invasive potential and neurosphere formation in vitro compared to EpCAM(–) Y79 cells. EpCAM(+) cells showed higher β-catenin expression compared to EpCAMˉ cells. EpCAM×CD3 significantly retarded proliferation of RB primary tumor cells. EpCAM×CD3 effectively induced the secretion of effector cytokines, such as interferon (IFN)-γ, tumor necrosis factor (TNF)-α, interleukin (IL)-10, IL-2, and transforming growth factor (TGF)-β1, and also perforin levels by pre-activated lymphocytes. CONCLUSIONS: EpCAM might be a novel cancer stem cell marker in RB. EpCAM×CD3 antibody redirecting T cells to attack RB tumor cells may prove effective in RB management. Further preclinical studies are needed to confirm the initial findings of our study. Molecular Vision 2012-02-01 /pmc/articles/PMC3275636/ /pubmed/22328825 Text en Copyright © 2012 Molecular Vision. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Mitra, Moutushy Kandalam, Mallikarjuna Harilal, Anju Verma, Rama Shenkar Krishnan, Uma Maheswari Swaminathan, Sethuraman Krishnakumar, Subramanian EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy |
title | EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy |
title_full | EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy |
title_fullStr | EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy |
title_full_unstemmed | EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy |
title_short | EpCAM is a putative stem marker in retinoblastoma and an effective target for T-cell-mediated immunotherapy |
title_sort | epcam is a putative stem marker in retinoblastoma and an effective target for t-cell-mediated immunotherapy |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3275636/ https://www.ncbi.nlm.nih.gov/pubmed/22328825 |
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