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Conjunctival melanoma and electrochemotherapy: preliminary results using 2D and 3D cell culture models in vitro

PURPOSE: To investigate the cytotoxic effect of bleomycin, mitomycin C (MMC) and Fluorouracil (5‐FU) in combination with electroporation (EP) on human conjunctival melanoma (CM) and normal conjunctival cell lines using 2D and 3D cell culture systems in vitro. METHODS: Two CM (CRMM1, CRMM2) and one n...

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Autores principales: Fiorentzis, Miltiadis, Katopodis, Periklis, Kalirai, Helen, Seitz, Berthold, Viestenz, Arne, Coupland, Sarah E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6590119/
https://www.ncbi.nlm.nih.gov/pubmed/30548215
http://dx.doi.org/10.1111/aos.13993
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author Fiorentzis, Miltiadis
Katopodis, Periklis
Kalirai, Helen
Seitz, Berthold
Viestenz, Arne
Coupland, Sarah E.
author_facet Fiorentzis, Miltiadis
Katopodis, Periklis
Kalirai, Helen
Seitz, Berthold
Viestenz, Arne
Coupland, Sarah E.
author_sort Fiorentzis, Miltiadis
collection PubMed
description PURPOSE: To investigate the cytotoxic effect of bleomycin, mitomycin C (MMC) and Fluorouracil (5‐FU) in combination with electroporation (EP) on human conjunctival melanoma (CM) and normal conjunctival cell lines using 2D and 3D cell culture systems in vitro. METHODS: Two CM (CRMM1, CRMM2) and one normal conjunctival epithelial cell line (HCjE‐Gi) were treated with various EP conditions and increasing concentrations of 5‐FU, MMC and bleomycin. Cell survival was assessed by MTT viability assay. All cell lines were seeded to create spheroids and were treated with bleomycin on day 3 and day 8 combined with EP. Spheroids were collected, fixed in buffered formalin and subsequently paraffin embedded for histological assessment of the effects of the treatment on cell viability. RESULTS: CM cell lines were resistant to electroporation alone and showed a reduction in cell number only when treated with 1000 Volts/cm and 8 pulses. HCjE‐Gi cells showed higher sensitivity to electric pulses over 750 Volts/cm. MMC and 5‐FU demonstrated a higher cytotoxicity for the HCjE‐Gi cell line. The CM cell lines were resistant to MMC and 5‐FU. Bleomycin (1 μg/ml) alone had no significant effect on the HCjE‐Gi even when combined with EP conditions ≥750 Volts/cm. In contrast, it significantly (p ‐, paired t‐test) reduced cell viability in the CM cell lines. Spheroids treated with bleomycin and EP showed a reduction in tumour mass and proliferation rates after treatment. CONCLUSION: Our in vitro study using 2D and 3D models indicates that the application of EP may effectively enhance chemotherapy with bleomycin in CM. This may offer new viable perspectives for CM treatment.
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spelling pubmed-65901192019-07-08 Conjunctival melanoma and electrochemotherapy: preliminary results using 2D and 3D cell culture models in vitro Fiorentzis, Miltiadis Katopodis, Periklis Kalirai, Helen Seitz, Berthold Viestenz, Arne Coupland, Sarah E. Acta Ophthalmol Original Articles PURPOSE: To investigate the cytotoxic effect of bleomycin, mitomycin C (MMC) and Fluorouracil (5‐FU) in combination with electroporation (EP) on human conjunctival melanoma (CM) and normal conjunctival cell lines using 2D and 3D cell culture systems in vitro. METHODS: Two CM (CRMM1, CRMM2) and one normal conjunctival epithelial cell line (HCjE‐Gi) were treated with various EP conditions and increasing concentrations of 5‐FU, MMC and bleomycin. Cell survival was assessed by MTT viability assay. All cell lines were seeded to create spheroids and were treated with bleomycin on day 3 and day 8 combined with EP. Spheroids were collected, fixed in buffered formalin and subsequently paraffin embedded for histological assessment of the effects of the treatment on cell viability. RESULTS: CM cell lines were resistant to electroporation alone and showed a reduction in cell number only when treated with 1000 Volts/cm and 8 pulses. HCjE‐Gi cells showed higher sensitivity to electric pulses over 750 Volts/cm. MMC and 5‐FU demonstrated a higher cytotoxicity for the HCjE‐Gi cell line. The CM cell lines were resistant to MMC and 5‐FU. Bleomycin (1 μg/ml) alone had no significant effect on the HCjE‐Gi even when combined with EP conditions ≥750 Volts/cm. In contrast, it significantly (p ‐, paired t‐test) reduced cell viability in the CM cell lines. Spheroids treated with bleomycin and EP showed a reduction in tumour mass and proliferation rates after treatment. CONCLUSION: Our in vitro study using 2D and 3D models indicates that the application of EP may effectively enhance chemotherapy with bleomycin in CM. This may offer new viable perspectives for CM treatment. John Wiley and Sons Inc. 2018-12-10 2019-06 /pmc/articles/PMC6590119/ /pubmed/30548215 http://dx.doi.org/10.1111/aos.13993 Text en © 2018 The Authors. Acta Ophthalmologica published by John Wiley & Sons Ltd on behalf of Acta Ophthalmologica Scandinavica Foundation. This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.
spellingShingle Original Articles
Fiorentzis, Miltiadis
Katopodis, Periklis
Kalirai, Helen
Seitz, Berthold
Viestenz, Arne
Coupland, Sarah E.
Conjunctival melanoma and electrochemotherapy: preliminary results using 2D and 3D cell culture models in vitro
title Conjunctival melanoma and electrochemotherapy: preliminary results using 2D and 3D cell culture models in vitro
title_full Conjunctival melanoma and electrochemotherapy: preliminary results using 2D and 3D cell culture models in vitro
title_fullStr Conjunctival melanoma and electrochemotherapy: preliminary results using 2D and 3D cell culture models in vitro
title_full_unstemmed Conjunctival melanoma and electrochemotherapy: preliminary results using 2D and 3D cell culture models in vitro
title_short Conjunctival melanoma and electrochemotherapy: preliminary results using 2D and 3D cell culture models in vitro
title_sort conjunctival melanoma and electrochemotherapy: preliminary results using 2d and 3d cell culture models in vitro
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6590119/
https://www.ncbi.nlm.nih.gov/pubmed/30548215
http://dx.doi.org/10.1111/aos.13993
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