Effects of N-acetyl-l-cysteine on adhesive strength between breast cancer cell and extracellular matrix proteins after ionizing radiation
Aims: To evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular matrix (ECM) proteins after IR. Main methods: Using static cell adhesion assays to determine the effect of various times and duration...
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| Published in | Life sciences (1973) Vol. 93; no. 21; pp. 798 - 803 |
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| Main Authors | , , |
| Format | Journal Article |
| Language | English |
| Published |
Netherlands
Elsevier Inc
13.11.2013
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| Subjects | |
| Online Access | Get full text |
| ISSN | 0024-3205 1879-0631 1879-0631 |
| DOI | 10.1016/j.lfs.2013.09.029 |
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| Abstract | Aims: To evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular matrix (ECM) proteins after IR.
Main methods: Using static cell adhesion assays to determine the effect of various times and duration of LNAC (10mM) treatment on IR (20Gy)-altered adhesive affinity between MDA-MB-231 breast cancer cells and ECM, especially fibronectin; using fluorescence dye carboxy- 2,7-dichlorodihydrofluorescein diacetate to determine intracellular levels of ROS; using flow cytometry to determine cell surface integrin β1; and using Western blot analysis to determine vimentin expression.
Key findings: Our results indicated that continuously treating the breast cancer cells with LNAC for 24h, starting immediately after IR, could inhibit IR-induced cell adhesion to ECM proteins at 24h post-IR. The reduction of cell adhesive affinity was correlated with a down-regulation of IR-induced ROS production and surface expression of activated integrin β1. When the cells were pretreated for 1h, the inhibitory effects of LNAC were found to be either reduced or completely abrogated followed by 24h or 2h treatments, respectively. In addition to cell adhesion, treatment with LNAC inhibited IR-induced expression of vimentin, an epithelial–mesenchymal transition marker (EMT).
Significance: The benefits of administering antioxidants during radiation therapy have been the subject of much controversy. Our results suggest that if antioxidant treatment is to be combined with IR therapy, time of administration and treatment duration are important variables to consider. |
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| AbstractList | Aims: To evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular matrix (ECM) proteins after IR. Main methods: Using static cell adhesion assays to determine the effect of various times and duration of LNAC (10mM) treatment on IR (20Gy)-altered adhesive affinity between MDA-MB-231 breast cancer cells and ECM, especially fibronectin; using fluorescence dye carboxy- 2,7-dichlorodihydrofluorescein diacetate to determine intracellular levels of ROS; using flow cytometry to determine cell surface integrin β1; and using Western blot analysis to determine vimentin expression. Key findings: Our results indicated that continuously treating the breast cancer cells with LNAC for 24h, starting immediately after IR, could inhibit IR-induced cell adhesion to ECM proteins at 24h post-IR. The reduction of cell adhesive affinity was correlated with a down-regulation of IR-induced ROS production and surface expression of activated integrin β1. When the cells were pretreated for 1h, the inhibitory effects of LNAC were found to be either reduced or completely abrogated followed by 24h or 2h treatments, respectively. In addition to cell adhesion, treatment with LNAC inhibited IR-induced expression of vimentin, an epithelial–mesenchymal transition marker (EMT). Significance: The benefits of administering antioxidants during radiation therapy have been the subject of much controversy. Our results suggest that if antioxidant treatment is to be combined with IR therapy, time of administration and treatment duration are important variables to consider. To evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular matrix (ECM) proteins after IR. Using static cell adhesion assays to determine the effect of various times and duration of LNAC (10mM) treatment on IR (20Gy)-altered adhesive affinity between MDA-MB-231 breast cancer cells and ECM, especially fibronectin; using fluorescence dye carboxy- 2,7-dichlorodihydrofluorescein diacetate to determine intracellular levels of ROS; using flow cytometry to determine cell surface integrin β1; and using Western blot analysis to determine vimentin expression. Our results indicated that continuously treating the breast cancer cells with LNAC for 24h, starting immediately after IR, could inhibit IR-induced cell adhesion to ECM proteins at 24h post-IR. The reduction of cell adhesive affinity was correlated with a down-regulation of IR-induced ROS production and surface expression of activated integrin β1. When the cells were pretreated for 1h, the inhibitory effects of LNAC were found to be either reduced or completely abrogated followed by 24h or 2h treatments, respectively. In addition to cell adhesion, treatment with LNAC inhibited IR-induced expression of vimentin, an epithelial-mesenchymal transition marker (EMT). The benefits of administering antioxidants during radiation therapy have been the subject of much controversy. Our results suggest that if antioxidant treatment is to be combined with IR therapy, time of administration and treatment duration are important variables to consider. The benefits of administering antioxidants during radiation therapy have been the subject of much controversy. It is known that both ionizing radiation (IR) and reactive oxygen species (ROS) alter the adhesive affinity between tumor cells and extracellular matrix (ECM) proteins and that IR induces production of ROS. We therefore evaluated the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular matrix (ECM) proteins after IR. We focused mainly on fibronectin, a representative ECM protein. Our results using static cell adhesion assays indicated that continuously treating the breast cancer cells with LNAC (10 mM) for 24 h, starting immediately after IR (20 Gy), could inhibit IR-induced cell adhesion to ECM proteins at 24 h post-IR. Analyses of intracellular levels of ROS by the fluorescence dye carboxy- 2,7-dichlorodihydrofluorescin diacetate and activated integrin β1 by flow cytometry revealed that the reduction of cell adhesive affinity was correlated with a down-regulation of IR-induced ROS production and surface expression of activated integrin β1. In addition to cell adhesion, treatment with LNAC inhibited IR-induced expression of vimentin, an epithelial mesenchymal transition marker (EMT). Interestingly, when the cells were pretreated for 1 h, the inhibitory effects of LNAC were found to be either reduced or completely abrogated followed by 24 h or 2 h treatments, respectively. Our results demonstrated that the time and duration of LNAC treatment is critical for regulating IR-induced adhesive affinity, and thus metastatic potential, as well as EMT process of breast cancer cells. Aims: To evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular matrix (ECM) proteins after IR. Main methods: Using static cell adhesion assays to determine the effect of various times and duration of LNAC (10mM) treatment on IR (20Gy)-altered adhesive affinity between MDA-MB-231 breast cancer cells and ECM, especially fibronectin; using fluorescence dye carboxy- 2,7-dichlorodihydrofluorescein diacetate to determine intracellular levels of ROS; using flow cytometry to determine cell surface integrin β1; and using Western blot analysis to determine vimentin expression. Key findings: Our results indicated that continuously treating the breast cancer cells with LNAC for 24h, starting immediately after IR, could inhibit IR-induced cell adhesion to ECM proteins at 24h post-IR. The reduction of cell adhesive affinity was correlated with a down-regulation of IR-induced ROS production and surface expression of activated integrin β1. When the cells were pretreated for 1h, the inhibitory effects of LNAC were found to be either reduced or completely abrogated followed by 24h or 2h treatments, respectively. In addition to cell adhesion, treatment with LNAC inhibited IR-induced expression of vimentin, an epithelial–mesenchymal transition marker (EMT). Significance: The benefits of administering antioxidants during radiation therapy have been the subject of much controversy. Our results suggest that if antioxidant treatment is to be combined with IR therapy, time of administration and treatment duration are important variables to consider. To evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular matrix (ECM) proteins after IR.AIMSTo evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular matrix (ECM) proteins after IR.Using static cell adhesion assays to determine the effect of various times and duration of LNAC (10mM) treatment on IR (20Gy)-altered adhesive affinity between MDA-MB-231 breast cancer cells and ECM, especially fibronectin; using fluorescence dye carboxy- 2,7-dichlorodihydrofluorescein diacetate to determine intracellular levels of ROS; using flow cytometry to determine cell surface integrin β1; and using Western blot analysis to determine vimentin expression.MAIN METHODSUsing static cell adhesion assays to determine the effect of various times and duration of LNAC (10mM) treatment on IR (20Gy)-altered adhesive affinity between MDA-MB-231 breast cancer cells and ECM, especially fibronectin; using fluorescence dye carboxy- 2,7-dichlorodihydrofluorescein diacetate to determine intracellular levels of ROS; using flow cytometry to determine cell surface integrin β1; and using Western blot analysis to determine vimentin expression.Our results indicated that continuously treating the breast cancer cells with LNAC for 24h, starting immediately after IR, could inhibit IR-induced cell adhesion to ECM proteins at 24h post-IR. The reduction of cell adhesive affinity was correlated with a down-regulation of IR-induced ROS production and surface expression of activated integrin β1. When the cells were pretreated for 1h, the inhibitory effects of LNAC were found to be either reduced or completely abrogated followed by 24h or 2h treatments, respectively. In addition to cell adhesion, treatment with LNAC inhibited IR-induced expression of vimentin, an epithelial-mesenchymal transition marker (EMT).KEY FINDINGSOur results indicated that continuously treating the breast cancer cells with LNAC for 24h, starting immediately after IR, could inhibit IR-induced cell adhesion to ECM proteins at 24h post-IR. The reduction of cell adhesive affinity was correlated with a down-regulation of IR-induced ROS production and surface expression of activated integrin β1. When the cells were pretreated for 1h, the inhibitory effects of LNAC were found to be either reduced or completely abrogated followed by 24h or 2h treatments, respectively. In addition to cell adhesion, treatment with LNAC inhibited IR-induced expression of vimentin, an epithelial-mesenchymal transition marker (EMT).The benefits of administering antioxidants during radiation therapy have been the subject of much controversy. Our results suggest that if antioxidant treatment is to be combined with IR therapy, time of administration and treatment duration are important variables to consider.SIGNIFICANCEThe benefits of administering antioxidants during radiation therapy have been the subject of much controversy. Our results suggest that if antioxidant treatment is to be combined with IR therapy, time of administration and treatment duration are important variables to consider. |
| Author | Cheng, Huiwen Lee, Shin Hee Wu, Shiyong |
| AuthorAffiliation | a Edison Biotechnology Institute, Ohio University, Athens, Ohio 45701, USA b Department of Chemistry and Biochemistry, Ohio University, Athens, Ohio 45701, USA |
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| Keywords | Ionizing radiation Breast cancer Integrin LNAC Cell adhesion |
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| Snippet | Aims: To evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and... To evaluate the effect of N-acetyl-L-cysteine (LNAC), a common ROS scavenger, on the adhesive affinity between MDA-MB-231 breast cancer cells and extracellular... The benefits of administering antioxidants during radiation therapy have been the subject of much controversy. It is known that both ionizing radiation (IR)... |
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| SubjectTerms | acetylcysteine Acetylcysteine - administration & dosage Acetylcysteine - pharmacology antioxidants Breast cancer breast neoplasms Breast Neoplasms - pathology Catalase - pharmacology CD29 antigen Cell adhesion Cell Adhesion - drug effects Cell Adhesion - radiation effects Down-Regulation - drug effects Drug Administration Schedule extracellular matrix Extracellular Matrix Proteins - metabolism fibronectins flow cytometry fluorescence Free Radical Scavengers - administration & dosage Free Radical Scavengers - pharmacology Humans Integrin Integrin beta1 - metabolism Ionizing radiation LNAC neoplasm cells Radiation, Ionizing radiotherapy Reactive Oxygen Species - metabolism Tumor Cells, Cultured vimentin Vimentin - metabolism Western blotting |
| Title | Effects of N-acetyl-l-cysteine on adhesive strength between breast cancer cell and extracellular matrix proteins after ionizing radiation |
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