Cargando…
Decrease of ABCB1 protein expression and increase of G(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells
Oleanolic acid (OA) is a natural compound that can be found in a number of edible and medicinal plants and confers diverse biological actions. However, the direct target of OA in human tumor cells remains poorly understood, preventing its application in clinical and health settings. A previous study...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
D.A. Spandidos
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8138263/ https://www.ncbi.nlm.nih.gov/pubmed/34055052 http://dx.doi.org/10.3892/etm.2021.10167 |
_version_ | 1783695768453906432 |
---|---|
author | Wang, Didi Wang, Jincai Zhang, Juan Yi, Xin Piao, Jinhua Li, Li Wang, Jianjie Zhang, Pengxia He, Qiyang |
author_facet | Wang, Didi Wang, Jincai Zhang, Juan Yi, Xin Piao, Jinhua Li, Li Wang, Jianjie Zhang, Pengxia He, Qiyang |
author_sort | Wang, Didi |
collection | PubMed |
description | Oleanolic acid (OA) is a natural compound that can be found in a number of edible and medicinal plants and confers diverse biological actions. However, the direct target of OA in human tumor cells remains poorly understood, preventing its application in clinical and health settings. A previous study revealed that overexpression of caveolin-1 in human leukemia HL-60 cells can increase its sensitivity to OA. The present study aimed to investigate the effects of OA on the doxorubicin-resistant human breast cancer MCF-7 cell line (MCF-7/DOX), harringtonine-resistant human leukemia HL-60 cells (HL-60/HAR) and their corresponding parental cell lines. Western blotting was performed to measure protein expression levels, whilst Cell Counting Kit-8 (CCK-8) assays, cell cycle analysis (by flow cytometry) and apoptosis assays (with Annexin V/PI staining) were used to assess drug sensitivity. CCK-8 assay results suggested that MCF-7/DOX cells, which overexpress the caveolin-1 protein, have similar OA susceptibility to their parent line. In addition, sensitivity of MCF-7/DOX cells to OA was not augmented by knocking down caveolin-1 using RNA interference. HL-60/HAR cells exhibited a four-fold increased sensitivity to OA compared with that in their parental HL-60 cells according to CCK-8 assay. Both of the resistant cell lines exhibited higher numbers of cells at G(1) phase arrest compared with those in their parent lines, as measured via flow cytometry. Treatment of both MCF-7 cell lines with 100 µM OA for 48 h induced apoptosis, with increased effects observed in resistant cells. However, no PARP-1 or caspase-3 cleavage was observed, with some positive Annexin V staining found after HL-60/HAR cells were treated with OA, suggesting that cell death occurred via non-classical apoptosis or through other cell death pathways. It was found that OA was not a substrate of ATP-binding cassette subfamily B member 1 (ABCB1) in drug-resistant cells, as indicated by the accumulation of rhodamine 123 assessed using flow cytometry. However, protein expression of ABCB1 in both of the resistant cell lines was significantly decreased after treatment with OA in a concentration-dependent manner. Collectively, these results suggest that OA could reduce ABCB1 protein expression and induce G(1) phase arrest in multidrug-resistant cancer cells. These findings highlight the potential of OA for cancer therapy. |
format | Online Article Text |
id | pubmed-8138263 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | D.A. Spandidos |
record_format | MEDLINE/PubMed |
spelling | pubmed-81382632021-05-27 Decrease of ABCB1 protein expression and increase of G(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells Wang, Didi Wang, Jincai Zhang, Juan Yi, Xin Piao, Jinhua Li, Li Wang, Jianjie Zhang, Pengxia He, Qiyang Exp Ther Med Articles Oleanolic acid (OA) is a natural compound that can be found in a number of edible and medicinal plants and confers diverse biological actions. However, the direct target of OA in human tumor cells remains poorly understood, preventing its application in clinical and health settings. A previous study revealed that overexpression of caveolin-1 in human leukemia HL-60 cells can increase its sensitivity to OA. The present study aimed to investigate the effects of OA on the doxorubicin-resistant human breast cancer MCF-7 cell line (MCF-7/DOX), harringtonine-resistant human leukemia HL-60 cells (HL-60/HAR) and their corresponding parental cell lines. Western blotting was performed to measure protein expression levels, whilst Cell Counting Kit-8 (CCK-8) assays, cell cycle analysis (by flow cytometry) and apoptosis assays (with Annexin V/PI staining) were used to assess drug sensitivity. CCK-8 assay results suggested that MCF-7/DOX cells, which overexpress the caveolin-1 protein, have similar OA susceptibility to their parent line. In addition, sensitivity of MCF-7/DOX cells to OA was not augmented by knocking down caveolin-1 using RNA interference. HL-60/HAR cells exhibited a four-fold increased sensitivity to OA compared with that in their parental HL-60 cells according to CCK-8 assay. Both of the resistant cell lines exhibited higher numbers of cells at G(1) phase arrest compared with those in their parent lines, as measured via flow cytometry. Treatment of both MCF-7 cell lines with 100 µM OA for 48 h induced apoptosis, with increased effects observed in resistant cells. However, no PARP-1 or caspase-3 cleavage was observed, with some positive Annexin V staining found after HL-60/HAR cells were treated with OA, suggesting that cell death occurred via non-classical apoptosis or through other cell death pathways. It was found that OA was not a substrate of ATP-binding cassette subfamily B member 1 (ABCB1) in drug-resistant cells, as indicated by the accumulation of rhodamine 123 assessed using flow cytometry. However, protein expression of ABCB1 in both of the resistant cell lines was significantly decreased after treatment with OA in a concentration-dependent manner. Collectively, these results suggest that OA could reduce ABCB1 protein expression and induce G(1) phase arrest in multidrug-resistant cancer cells. These findings highlight the potential of OA for cancer therapy. D.A. Spandidos 2021-07 2021-05-09 /pmc/articles/PMC8138263/ /pubmed/34055052 http://dx.doi.org/10.3892/etm.2021.10167 Text en Copyright: © Wang et al. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , 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 | Articles Wang, Didi Wang, Jincai Zhang, Juan Yi, Xin Piao, Jinhua Li, Li Wang, Jianjie Zhang, Pengxia He, Qiyang Decrease of ABCB1 protein expression and increase of G(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells |
title | Decrease of ABCB1 protein expression and increase of G(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells |
title_full | Decrease of ABCB1 protein expression and increase of G(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells |
title_fullStr | Decrease of ABCB1 protein expression and increase of G(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells |
title_full_unstemmed | Decrease of ABCB1 protein expression and increase of G(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells |
title_short | Decrease of ABCB1 protein expression and increase of G(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells |
title_sort | decrease of abcb1 protein expression and increase of g(1) phase arrest induced by oleanolic acid in human multidrug-resistant cancer cells |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8138263/ https://www.ncbi.nlm.nih.gov/pubmed/34055052 http://dx.doi.org/10.3892/etm.2021.10167 |
work_keys_str_mv | AT wangdidi decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells AT wangjincai decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells AT zhangjuan decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells AT yixin decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells AT piaojinhua decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells AT lili decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells AT wangjianjie decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells AT zhangpengxia decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells AT heqiyang decreaseofabcb1proteinexpressionandincreaseofg1phasearrestinducedbyoleanolicacidinhumanmultidrugresistantcancercells |