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Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells
BACKGROUND: Ursolic acid (UA) is an anti-cancer herbal compound. In the present study, we observed the effects of UA on anchorage-dependent and -independent growth of human colorectal cancer (CRC) RKO cells. METHODS: RKO cells were cultured in conventional and detached condition and treated with UA....
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7866452/ https://www.ncbi.nlm.nih.gov/pubmed/33549076 http://dx.doi.org/10.1186/s12906-021-03232-2 |
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author | Zheng, Jia-Lu Wang, Shuang-Shuang Shen, Ke-Ping Chen, Lei Peng, Xiao Chen, Jin-Fang An, Hong-Mei Hu, Bing |
author_facet | Zheng, Jia-Lu Wang, Shuang-Shuang Shen, Ke-Ping Chen, Lei Peng, Xiao Chen, Jin-Fang An, Hong-Mei Hu, Bing |
author_sort | Zheng, Jia-Lu |
collection | PubMed |
description | BACKGROUND: Ursolic acid (UA) is an anti-cancer herbal compound. In the present study, we observed the effects of UA on anchorage-dependent and -independent growth of human colorectal cancer (CRC) RKO cells. METHODS: RKO cells were cultured in conventional and detached condition and treated with UA. Cell viability was evaluated by CCK-8 assay. Cell cycle was analyzed by flow cytometry. Apoptosis was identified by Hoechst 33258 staining and flow cytometry analysis. Activities of caspases were measured by commercial kits. Reactive oxygen species (ROS) was recognized by DCFH-DA fluorescent staining. Anoikis was identified by EthD-1 fluorescent staining and flow cytometry analysis. Expression and phosphorylation of proteins were analyzed by western blot. RESULTS: UA inhibited RKO cell viability in both a dose- and time-dependent manner. UA arrested the cell cycle at the G0/G1 phase, and induced caspase-dependent apoptosis. UA inhibited Bcl-2 expression and increased Bax expression. In addition, UA up-regulated the level of ROS that contributed to UA activated caspase-3, − 8 and − 9, and induced apoptosis. Furthermore, UA inhibited cell growth in a detached condition and induced anoikis in RKO cells that was accompanied by dampened phosphorylation of FAK, PI3K and AKT. UA also inhibited epithelial-mesenchymal transition (EMT) as indicated by the down-regulation of N-Cad expression and up-regulation of E-Cad expression. CONCLUSIONS: UA induced caspase-dependent apoptosis, and FAK/PI3K/AKT singling and EMT related anoikis in RKO cells. UA was an effective anti-cancer compound against both anchorage-dependent and -independent growth of RKO cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12906-021-03232-2. |
format | Online Article Text |
id | pubmed-7866452 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-78664522021-02-08 Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells Zheng, Jia-Lu Wang, Shuang-Shuang Shen, Ke-Ping Chen, Lei Peng, Xiao Chen, Jin-Fang An, Hong-Mei Hu, Bing BMC Complement Med Ther Research Article BACKGROUND: Ursolic acid (UA) is an anti-cancer herbal compound. In the present study, we observed the effects of UA on anchorage-dependent and -independent growth of human colorectal cancer (CRC) RKO cells. METHODS: RKO cells were cultured in conventional and detached condition and treated with UA. Cell viability was evaluated by CCK-8 assay. Cell cycle was analyzed by flow cytometry. Apoptosis was identified by Hoechst 33258 staining and flow cytometry analysis. Activities of caspases were measured by commercial kits. Reactive oxygen species (ROS) was recognized by DCFH-DA fluorescent staining. Anoikis was identified by EthD-1 fluorescent staining and flow cytometry analysis. Expression and phosphorylation of proteins were analyzed by western blot. RESULTS: UA inhibited RKO cell viability in both a dose- and time-dependent manner. UA arrested the cell cycle at the G0/G1 phase, and induced caspase-dependent apoptosis. UA inhibited Bcl-2 expression and increased Bax expression. In addition, UA up-regulated the level of ROS that contributed to UA activated caspase-3, − 8 and − 9, and induced apoptosis. Furthermore, UA inhibited cell growth in a detached condition and induced anoikis in RKO cells that was accompanied by dampened phosphorylation of FAK, PI3K and AKT. UA also inhibited epithelial-mesenchymal transition (EMT) as indicated by the down-regulation of N-Cad expression and up-regulation of E-Cad expression. CONCLUSIONS: UA induced caspase-dependent apoptosis, and FAK/PI3K/AKT singling and EMT related anoikis in RKO cells. UA was an effective anti-cancer compound against both anchorage-dependent and -independent growth of RKO cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12906-021-03232-2. BioMed Central 2021-02-06 /pmc/articles/PMC7866452/ /pubmed/33549076 http://dx.doi.org/10.1186/s12906-021-03232-2 Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Article Zheng, Jia-Lu Wang, Shuang-Shuang Shen, Ke-Ping Chen, Lei Peng, Xiao Chen, Jin-Fang An, Hong-Mei Hu, Bing Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells |
title | Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells |
title_full | Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells |
title_fullStr | Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells |
title_full_unstemmed | Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells |
title_short | Ursolic acid induces apoptosis and anoikis in colorectal carcinoma RKO cells |
title_sort | ursolic acid induces apoptosis and anoikis in colorectal carcinoma rko cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7866452/ https://www.ncbi.nlm.nih.gov/pubmed/33549076 http://dx.doi.org/10.1186/s12906-021-03232-2 |
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