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Potential role of p53 on metallothionein induction in human epithelial breast cancer cells
The expression and induction of metallothionein has been associated with protection against oxidative stress and apoptosis. This study examines the effect of tumour suppressor protein p53 on metallothionein expression following CdCl(2) treatment in eight human epithelial breast cancer cell lines dif...
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Formato: | Texto |
Lenguaje: | English |
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Nature Publishing Group
2002
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2364318/ https://www.ncbi.nlm.nih.gov/pubmed/12434295 http://dx.doi.org/10.1038/sj.bjc.6600549 |
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author | Fan, L Z Cherian, M G |
author_facet | Fan, L Z Cherian, M G |
author_sort | Fan, L Z |
collection | PubMed |
description | The expression and induction of metallothionein has been associated with protection against oxidative stress and apoptosis. This study examines the effect of tumour suppressor protein p53 on metallothionein expression following CdCl(2) treatment in eight human epithelial breast cancer cell lines differing in p53 and oestrogen-receptor status. Cells were treated with 10 μM CdCl(2) for 24 h and metallothionein protein levels were measured by cadmium binding assay. MCF7 cells which are p53-positive (p53+) and oestrogen-receptor-positive showed a large induction in metallothionein synthesis by 10.79±1.36-fold. Other breast cancer cell lines which are p53-negative (p53−) and oestrogen-receptor-negative or weakly oestrogen-receptor-positive showed a small induction ranging from 1.40±0.10 to 3.65±0.30-fold. RT–PCR analysis showed an induction of metallothionein mRNA in MCF7 cells by about 1.61±0.08-fold, while in HCC1806 cells (p53−, oestrogen-receptor-negative) by 1.11±0.13-fold, and in MDA-MB-231 (p53−, oestrogen-receptor-negative) by 1.25±0.06-fold. Metallothionein localisation was determined by immunohistochemical staining. Prior to metal treatment, metallothionein was localised mainly in the cytoplasm of MCF7 and MDA-MB-231 cells. After treatment with 10 μM CdCl(2) for 24 h, MCF7 cells showed intense nuclear and cytoplasmic staining for metallothionein, while MDA-MB-231 cells showed staining in the cytoplasm with weak nuclear staining. Apoptosis induced by 10–40 μM CdCl(2) at time points between 4 and 48 h was examined with TUNEL assay. In MCF7 cells, apoptosis increased with higher concentrations of CdCl(2), it peaked at 6–8 h and appeared again at 48 h for all concentrations of CdCl(2) tested. In MDA-MB-231 cells, apoptosis remained at low levels for 10–40 μM CdCl(2) at all time points. Studies on cadmium uptake showed similar uptake and accumulation of cadmium at 8 and 24 h in all the cell lines. The data demonstrate that treatment of epithelial breast cancer cells with 10 μM CdCl(2) for 24 h caused a greater induction of metallothionein protein and mRNA expression in p53+ and oestrogen-receptor-positive cells as compared to p53− and oestrogen-receptor-negative or weakly oestrogen-receptor-positive cells. This effect may be associated with the occurrence of apoptosis and suggests a role for p53 and oestrogen-receptor on the expression and induction of metallothionein in epithelial cells. British Journal of Cancer (2002) 87, 1019–1026. doi:10.1038/sj.bjc.6600549 www.bjcancer.com © 2002 Cancer Research UK |
format | Text |
id | pubmed-2364318 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2002 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-23643182009-09-10 Potential role of p53 on metallothionein induction in human epithelial breast cancer cells Fan, L Z Cherian, M G Br J Cancer Experimental Therapeutics The expression and induction of metallothionein has been associated with protection against oxidative stress and apoptosis. This study examines the effect of tumour suppressor protein p53 on metallothionein expression following CdCl(2) treatment in eight human epithelial breast cancer cell lines differing in p53 and oestrogen-receptor status. Cells were treated with 10 μM CdCl(2) for 24 h and metallothionein protein levels were measured by cadmium binding assay. MCF7 cells which are p53-positive (p53+) and oestrogen-receptor-positive showed a large induction in metallothionein synthesis by 10.79±1.36-fold. Other breast cancer cell lines which are p53-negative (p53−) and oestrogen-receptor-negative or weakly oestrogen-receptor-positive showed a small induction ranging from 1.40±0.10 to 3.65±0.30-fold. RT–PCR analysis showed an induction of metallothionein mRNA in MCF7 cells by about 1.61±0.08-fold, while in HCC1806 cells (p53−, oestrogen-receptor-negative) by 1.11±0.13-fold, and in MDA-MB-231 (p53−, oestrogen-receptor-negative) by 1.25±0.06-fold. Metallothionein localisation was determined by immunohistochemical staining. Prior to metal treatment, metallothionein was localised mainly in the cytoplasm of MCF7 and MDA-MB-231 cells. After treatment with 10 μM CdCl(2) for 24 h, MCF7 cells showed intense nuclear and cytoplasmic staining for metallothionein, while MDA-MB-231 cells showed staining in the cytoplasm with weak nuclear staining. Apoptosis induced by 10–40 μM CdCl(2) at time points between 4 and 48 h was examined with TUNEL assay. In MCF7 cells, apoptosis increased with higher concentrations of CdCl(2), it peaked at 6–8 h and appeared again at 48 h for all concentrations of CdCl(2) tested. In MDA-MB-231 cells, apoptosis remained at low levels for 10–40 μM CdCl(2) at all time points. Studies on cadmium uptake showed similar uptake and accumulation of cadmium at 8 and 24 h in all the cell lines. The data demonstrate that treatment of epithelial breast cancer cells with 10 μM CdCl(2) for 24 h caused a greater induction of metallothionein protein and mRNA expression in p53+ and oestrogen-receptor-positive cells as compared to p53− and oestrogen-receptor-negative or weakly oestrogen-receptor-positive cells. This effect may be associated with the occurrence of apoptosis and suggests a role for p53 and oestrogen-receptor on the expression and induction of metallothionein in epithelial cells. British Journal of Cancer (2002) 87, 1019–1026. doi:10.1038/sj.bjc.6600549 www.bjcancer.com © 2002 Cancer Research UK Nature Publishing Group 2002-10-21 2002-10-21 /pmc/articles/PMC2364318/ /pubmed/12434295 http://dx.doi.org/10.1038/sj.bjc.6600549 Text en Copyright © 2002 Cancer Research UK https://creativecommons.org/licenses/by/4.0/This 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 license, and indicate if changes were made.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material.If material is not included in the article’s Creative Commons license 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 license, visit https://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Experimental Therapeutics Fan, L Z Cherian, M G Potential role of p53 on metallothionein induction in human epithelial breast cancer cells |
title | Potential role of p53 on metallothionein induction in human epithelial breast cancer cells |
title_full | Potential role of p53 on metallothionein induction in human epithelial breast cancer cells |
title_fullStr | Potential role of p53 on metallothionein induction in human epithelial breast cancer cells |
title_full_unstemmed | Potential role of p53 on metallothionein induction in human epithelial breast cancer cells |
title_short | Potential role of p53 on metallothionein induction in human epithelial breast cancer cells |
title_sort | potential role of p53 on metallothionein induction in human epithelial breast cancer cells |
topic | Experimental Therapeutics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2364318/ https://www.ncbi.nlm.nih.gov/pubmed/12434295 http://dx.doi.org/10.1038/sj.bjc.6600549 |
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