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Establishment and Characterization of a Novel Human Ocular Adnexal Sebaceous Carcinoma Cell Line

PURPOSE: Sebaceous carcinoma (SC) is a malignant eyelid tumor of the ocular adnexa that is primarily treated via surgical excision. Few therapies exist in advanced cases, and medical therapy is limited because of our incomplete understanding of SC biology. Herein, we describe a technique to culture...

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Autores principales: Rong, Andrew J., Gallo, Ryan A., Zhang, Michelle G., Doddapaneni, Ravi, Griswold, Anthony J., Lee, John Y., Kurtenbach, Stefan, Dubovy, Sander R., Tse, David T., Pelaez, Daniel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Association for Research in Vision and Ophthalmology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8161695/
https://www.ncbi.nlm.nih.gov/pubmed/34043754
http://dx.doi.org/10.1167/tvst.10.6.34
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author Rong, Andrew J.
Gallo, Ryan A.
Zhang, Michelle G.
Doddapaneni, Ravi
Griswold, Anthony J.
Lee, John Y.
Kurtenbach, Stefan
Dubovy, Sander R.
Tse, David T.
Pelaez, Daniel
author_facet Rong, Andrew J.
Gallo, Ryan A.
Zhang, Michelle G.
Doddapaneni, Ravi
Griswold, Anthony J.
Lee, John Y.
Kurtenbach, Stefan
Dubovy, Sander R.
Tse, David T.
Pelaez, Daniel
author_sort Rong, Andrew J.
collection PubMed
description PURPOSE: Sebaceous carcinoma (SC) is a malignant eyelid tumor of the ocular adnexa that is primarily treated via surgical excision. Few therapies exist in advanced cases, and medical therapy is limited because of our incomplete understanding of SC biology. Herein, we describe a technique to culture human ocular adnexal SC for use as an in vitro model. METHODS: Human ocular adnexal SC tumor cells were isolated from a patient undergoing orbital exenteration surgery and named Bascom Palmer 50 (BP50). They were cultured in Dulbecco's modified Eagle medium/nutrient mixture F-12 supplemented with 10% fetal bovine serum and antibiotics and were maintained at 37°C in humidified 5% CO(2). The cells were characterized by immunohistochemistry, exome sequencing, and short tandem repeats analysis. In vitro drug screening against mitomycin-C (MMC) was performed using a cell viability assay. RESULTS: BP50 grew past 40 passages with a doubling time of 52.3 hours. Immunocytochemical staining revealed expression of SC-associated markers adipophilin, epithelial membrane antigen, p53, and androgen receptor. Whole exome sequencing showed a significant carryover in somatic mutations between the tumor tissue and corresponding cell line, revealing genetic markers consistent with SC. MMC affected cell viability in a dose-dependent manner. CONCLUSIONS: BP50 displays characteristics of ocular adnexal SC and therefore may facilitate improved understanding of SC biology and the high throughput assessment of novel therapeutic compounds and new drug combinatorial approaches targeted for this disease. TRANSLATIONAL RELEVANCE: Drug screening with MMC against these cells shows in vitro evidence to support its continued clinical use in SC.
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spelling pubmed-81616952021-06-09 Establishment and Characterization of a Novel Human Ocular Adnexal Sebaceous Carcinoma Cell Line Rong, Andrew J. Gallo, Ryan A. Zhang, Michelle G. Doddapaneni, Ravi Griswold, Anthony J. Lee, John Y. Kurtenbach, Stefan Dubovy, Sander R. Tse, David T. Pelaez, Daniel Transl Vis Sci Technol Methods PURPOSE: Sebaceous carcinoma (SC) is a malignant eyelid tumor of the ocular adnexa that is primarily treated via surgical excision. Few therapies exist in advanced cases, and medical therapy is limited because of our incomplete understanding of SC biology. Herein, we describe a technique to culture human ocular adnexal SC for use as an in vitro model. METHODS: Human ocular adnexal SC tumor cells were isolated from a patient undergoing orbital exenteration surgery and named Bascom Palmer 50 (BP50). They were cultured in Dulbecco's modified Eagle medium/nutrient mixture F-12 supplemented with 10% fetal bovine serum and antibiotics and were maintained at 37°C in humidified 5% CO(2). The cells were characterized by immunohistochemistry, exome sequencing, and short tandem repeats analysis. In vitro drug screening against mitomycin-C (MMC) was performed using a cell viability assay. RESULTS: BP50 grew past 40 passages with a doubling time of 52.3 hours. Immunocytochemical staining revealed expression of SC-associated markers adipophilin, epithelial membrane antigen, p53, and androgen receptor. Whole exome sequencing showed a significant carryover in somatic mutations between the tumor tissue and corresponding cell line, revealing genetic markers consistent with SC. MMC affected cell viability in a dose-dependent manner. CONCLUSIONS: BP50 displays characteristics of ocular adnexal SC and therefore may facilitate improved understanding of SC biology and the high throughput assessment of novel therapeutic compounds and new drug combinatorial approaches targeted for this disease. TRANSLATIONAL RELEVANCE: Drug screening with MMC against these cells shows in vitro evidence to support its continued clinical use in SC. The Association for Research in Vision and Ophthalmology 2021-05-27 /pmc/articles/PMC8161695/ /pubmed/34043754 http://dx.doi.org/10.1167/tvst.10.6.34 Text en Copyright 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
spellingShingle Methods
Rong, Andrew J.
Gallo, Ryan A.
Zhang, Michelle G.
Doddapaneni, Ravi
Griswold, Anthony J.
Lee, John Y.
Kurtenbach, Stefan
Dubovy, Sander R.
Tse, David T.
Pelaez, Daniel
Establishment and Characterization of a Novel Human Ocular Adnexal Sebaceous Carcinoma Cell Line
title Establishment and Characterization of a Novel Human Ocular Adnexal Sebaceous Carcinoma Cell Line
title_full Establishment and Characterization of a Novel Human Ocular Adnexal Sebaceous Carcinoma Cell Line
title_fullStr Establishment and Characterization of a Novel Human Ocular Adnexal Sebaceous Carcinoma Cell Line
title_full_unstemmed Establishment and Characterization of a Novel Human Ocular Adnexal Sebaceous Carcinoma Cell Line
title_short Establishment and Characterization of a Novel Human Ocular Adnexal Sebaceous Carcinoma Cell Line
title_sort establishment and characterization of a novel human ocular adnexal sebaceous carcinoma cell line
topic Methods
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8161695/
https://www.ncbi.nlm.nih.gov/pubmed/34043754
http://dx.doi.org/10.1167/tvst.10.6.34
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