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In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells

Jasione montana L. (Campanulaceae) is used in traditional Belarusian herbal medicine for sleep disorders in children, but the chemical composition and biological activity have not been investigated. In this study, the activities of J. montana extracts, their fractions and main compounds were evaluat...

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Autores principales: Juszczak, Aleksandra Maria, Czarnomysy, Robert, Strawa, Jakub Władysław, Zovko Končić, Marijana, Bielawski, Krzysztof, Tomczyk, Michał
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036727/
https://www.ncbi.nlm.nih.gov/pubmed/33805898
http://dx.doi.org/10.3390/ijms22073345
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author Juszczak, Aleksandra Maria
Czarnomysy, Robert
Strawa, Jakub Władysław
Zovko Končić, Marijana
Bielawski, Krzysztof
Tomczyk, Michał
author_facet Juszczak, Aleksandra Maria
Czarnomysy, Robert
Strawa, Jakub Władysław
Zovko Končić, Marijana
Bielawski, Krzysztof
Tomczyk, Michał
author_sort Juszczak, Aleksandra Maria
collection PubMed
description Jasione montana L. (Campanulaceae) is used in traditional Belarusian herbal medicine for sleep disorders in children, but the chemical composition and biological activity have not been investigated. In this study, the activities of J. montana extracts, their fractions and main compounds were evaluated in amelanotic melanoma C32 (CRL-1585) cells and normal fibroblasts (PCS-201-012). The extracts and fractions were analyzed using liquid chromatography–photodiode array detection–electrospray ionization–mass spectrometry (LC–PDA–ESI–MS/TOF) to characterize 25 compounds. Further, three major and known constituents, luteolin (22) and its derivatives such as 7-O-glucoside (12) and 7-O-sambubioside (9) were isolated and identified. The cytotoxic activities against fibroblasts and the amelanotic melanoma cell line were determined using the fixable viability stain (FVS) assay. The influence of diethyl ether (Et(2)O) fraction (JM4) and 22 on apoptosis induction was investigated using an annexin V binding assay. The obtained results showed significant cytotoxicity of JM4 and 22 with IC(50) values of 119.7 ± 3.2 and 95.1 ± 7.2 μg/mL, respectively. The proapoptotic potential after 22 treatment in the C32 human amelanotic melanoma cell line was comparable to that of vinblastine sulfate (VLB), detecting 29.2 ± 3.0% apoptotic cells. Moreover, 22 displayed less necrotic potential against melanoma cells than VLB. In addition, the influences of JM4 and 22 on the dysfunction of the mitochondrial membrane potential (MMP), cell cycle and activity of caspases 3, 8, 9, and 10 were established. The effects of JM4 on MMP change (74.5 ± 3.0% of the cells showed a reduced MMP) corresponded to the results obtained from the annexin V binding assay and activation of caspase-9. JM4 and 22 displayed a significant impact on caspase-9 (40.9 ± 2.4% of the cells contained active caspase-9 after JM4 treatment and 16.6 ± 0.8% after incubation with 22) and the intrinsic (mitochondrial) apoptotic pathway. Moreover, studies have shown that JM4 and 22 affect the activation of external apoptosis pathways by inducing the caspase-8 and caspase-10 cascades. Thus, activation of caspase-3 and DNA damage via external and internal apoptotic pathways were observed after treatment with JM4 and 22. The obtained results suggest that J. montana extracts could be developed as new topical preparations with potential anticancer properties due to their promising cytotoxic and proapoptotic potential.
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spelling pubmed-80367272021-04-12 In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells Juszczak, Aleksandra Maria Czarnomysy, Robert Strawa, Jakub Władysław Zovko Končić, Marijana Bielawski, Krzysztof Tomczyk, Michał Int J Mol Sci Article Jasione montana L. (Campanulaceae) is used in traditional Belarusian herbal medicine for sleep disorders in children, but the chemical composition and biological activity have not been investigated. In this study, the activities of J. montana extracts, their fractions and main compounds were evaluated in amelanotic melanoma C32 (CRL-1585) cells and normal fibroblasts (PCS-201-012). The extracts and fractions were analyzed using liquid chromatography–photodiode array detection–electrospray ionization–mass spectrometry (LC–PDA–ESI–MS/TOF) to characterize 25 compounds. Further, three major and known constituents, luteolin (22) and its derivatives such as 7-O-glucoside (12) and 7-O-sambubioside (9) were isolated and identified. The cytotoxic activities against fibroblasts and the amelanotic melanoma cell line were determined using the fixable viability stain (FVS) assay. The influence of diethyl ether (Et(2)O) fraction (JM4) and 22 on apoptosis induction was investigated using an annexin V binding assay. The obtained results showed significant cytotoxicity of JM4 and 22 with IC(50) values of 119.7 ± 3.2 and 95.1 ± 7.2 μg/mL, respectively. The proapoptotic potential after 22 treatment in the C32 human amelanotic melanoma cell line was comparable to that of vinblastine sulfate (VLB), detecting 29.2 ± 3.0% apoptotic cells. Moreover, 22 displayed less necrotic potential against melanoma cells than VLB. In addition, the influences of JM4 and 22 on the dysfunction of the mitochondrial membrane potential (MMP), cell cycle and activity of caspases 3, 8, 9, and 10 were established. The effects of JM4 on MMP change (74.5 ± 3.0% of the cells showed a reduced MMP) corresponded to the results obtained from the annexin V binding assay and activation of caspase-9. JM4 and 22 displayed a significant impact on caspase-9 (40.9 ± 2.4% of the cells contained active caspase-9 after JM4 treatment and 16.6 ± 0.8% after incubation with 22) and the intrinsic (mitochondrial) apoptotic pathway. Moreover, studies have shown that JM4 and 22 affect the activation of external apoptosis pathways by inducing the caspase-8 and caspase-10 cascades. Thus, activation of caspase-3 and DNA damage via external and internal apoptotic pathways were observed after treatment with JM4 and 22. The obtained results suggest that J. montana extracts could be developed as new topical preparations with potential anticancer properties due to their promising cytotoxic and proapoptotic potential. MDPI 2021-03-25 /pmc/articles/PMC8036727/ /pubmed/33805898 http://dx.doi.org/10.3390/ijms22073345 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Juszczak, Aleksandra Maria
Czarnomysy, Robert
Strawa, Jakub Władysław
Zovko Končić, Marijana
Bielawski, Krzysztof
Tomczyk, Michał
In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells
title In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells
title_full In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells
title_fullStr In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells
title_full_unstemmed In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells
title_short In Vitro Anticancer Potential of Jasione montana and Its Main Components against Human Amelanotic Melanoma Cells
title_sort in vitro anticancer potential of jasione montana and its main components against human amelanotic melanoma cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036727/
https://www.ncbi.nlm.nih.gov/pubmed/33805898
http://dx.doi.org/10.3390/ijms22073345
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