Decrease of cyclin D1 in the human lung adenocarcinoma cell line A-427 by 7-hydroxycoumarin
Coumarin in vivo has antitumor activity in various types of cancer. In vitro, coumarin and 7-hydroxycoumarin, its major biotransformation product in humans, inhibit the proliferation of several human tumor cell lines. The molecular mechanisms of these effects are unknown. To gain information about t...
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Published in | Lung cancer (Amsterdam, Netherlands) Vol. 34; no. 2; pp. 185 - 194 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
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Elsevier Ireland Ltd
01.11.2001
Elsevier Science |
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Abstract | Coumarin in vivo has antitumor activity in various types of cancer. In vitro, coumarin and 7-hydroxycoumarin, its major biotransformation product in humans, inhibit the proliferation of several human tumor cell lines. The molecular mechanisms of these effects are unknown. To gain information about these mechanisms, we studied the effects of coumarin and 7-hydroxycoumarin in the human lung adenocarcinoma cell line A-427 on the inhibition of: (i) cell proliferation; (ii) cell cycle progression; and (iii) expression of cyclins D1, E and A. The inhibitory concentrations 50 (IC
50) of both compounds were estimated by cytostatic assays of tetrazolium (MTT) reduction. The effects on cell cycle progression were assayed with propidium iodide and BrdU using DNA histograms and multiparametric flow cytometry. The percentages of cells expressing cyclins D1, E, and A were estimated by means of bivariate flow cytometry using propidium iodide, and FITC-conjugated monoclonal antibodies for each cyclin. The IC
50 (±S.E.M.
n=3) of 7-hydroxycoumarin and coumarin at 72 h exposure, were 100±4.8 and 257±8.8 μg/ml, respectively. 7-Hydroxycoumarin at the concentration of 160 μg/ml (1 mM), inhibited the G
1/S transition of the cell cycle, an action consistent with the cytostatic effect. No significant decreases of cyclins E and A were observed. In contrast, cyclin D1 significantly decreased, which appears to indicate an action of 7-hydroxycoumarin in early events of phase G
1. However, messenger RNA of cyclin D1, assayed by RT-PCR, did not change. This suggests a posttranscriptional effect. The effects of coumarin were not significant. Cyclin D1 is overexpressed in many types of cancer, and its inhibition has been proposed as a pharmacological and therapeutic target for novel antitumor agents. Knowledge of the decrease of cyclin D1 by 7-hydroxycoumarin may lead to its use in cancer therapy, as well as to the development of more active compounds. |
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AbstractList | Coumarin in vivo has antitumor activity in various types of cancer. In vitro, coumarin and 7-hydroxycoumarin, its major biotransformation product in humans, inhibit the proliferation of several human tumor cell lines. The molecular mechanisms of these effects are unknown. To gain information about these mechanisms, we studied the effects of coumarin and 7-hydroxycoumarin in the human lung adenocarcinoma cell line A-427 on the inhibition of: (i) cell proliferation; (ii) cell cycle progression; and (iii) expression of cyclins D1, E and A. The inhibitory concentrations 50 (IC
50) of both compounds were estimated by cytostatic assays of tetrazolium (MTT) reduction. The effects on cell cycle progression were assayed with propidium iodide and BrdU using DNA histograms and multiparametric flow cytometry. The percentages of cells expressing cyclins D1, E, and A were estimated by means of bivariate flow cytometry using propidium iodide, and FITC-conjugated monoclonal antibodies for each cyclin. The IC
50 (±S.E.M.
n=3) of 7-hydroxycoumarin and coumarin at 72 h exposure, were 100±4.8 and 257±8.8 μg/ml, respectively. 7-Hydroxycoumarin at the concentration of 160 μg/ml (1 mM), inhibited the G
1/S transition of the cell cycle, an action consistent with the cytostatic effect. No significant decreases of cyclins E and A were observed. In contrast, cyclin D1 significantly decreased, which appears to indicate an action of 7-hydroxycoumarin in early events of phase G
1. However, messenger RNA of cyclin D1, assayed by RT-PCR, did not change. This suggests a posttranscriptional effect. The effects of coumarin were not significant. Cyclin D1 is overexpressed in many types of cancer, and its inhibition has been proposed as a pharmacological and therapeutic target for novel antitumor agents. Knowledge of the decrease of cyclin D1 by 7-hydroxycoumarin may lead to its use in cancer therapy, as well as to the development of more active compounds. Coumarin in vivo has antitumor activity in various types of cancer. In vitro, coumarin and 7-hydroxycoumarin, its major biotransformation product in humans, inhibit the proliferation of several human tumor cell lines. The molecular mechanisms of these effects are unknown. To gain information about these mechanisms, we studied the effects of coumarin and 7-hydroxycoumarin in the human lung adenocarcinoma cell line A-427 on the inhibition of: (i) cell proliferation; (ii) cell cycle progression; and (iii) expression of cyclins D1, E and A. The inhibitory concentrations 50 (IC(50)) of both compounds were estimated by cytostatic assays of tetrazolium (MTT) reduction. The effects on cell cycle progression were assayed with propidium iodide and BrdU using DNA histograms and multiparametric flow cytometry. The percentages of cells expressing cyclins D1, E, and A were estimated by means of bivariate flow cytometry using propidium iodide, and FITC-conjugated monoclonal antibodies for each cyclin. The IC(50) (+/-S.E.M. n=3) of 7-hydroxycoumarin and coumarin at 72 h exposure, were 100+/-4.8 and 257+/-8.8 microg/ml, respectively. 7-Hydroxycoumarin at the concentration of 160 microg/ml (1 mM), inhibited the G(1)/S transition of the cell cycle, an action consistent with the cytostatic effect. No significant decreases of cyclins E and A were observed. In contrast, cyclin D1 significantly decreased, which appears to indicate an action of 7-hydroxycoumarin in early events of phase G(1). However, messenger RNA of cyclin D1, assayed by RT-PCR, did not change. This suggests a posttranscriptional effect. The effects of coumarin were not significant. Cyclin D1 is overexpressed in many types of cancer, and its inhibition has been proposed as a pharmacological and therapeutic target for novel antitumor agents. Knowledge of the decrease of cyclin D1 by 7-hydroxycoumarin may lead to its use in cancer therapy, as well as to the development of more active compounds. |
Author | Nieto-Rodriguez, Alejandro Velasco-Velázquez, Marco Antonio López-González, José Sullivan Garcı́a-Mondragón, Maria Juana Mendoza-Patiño, Nicandro Elizalde-Galvan, Patricia Molina-Guarneros, Juan Arcadio Jiménez-Orozco, Fausto Alejandro León-Cedeño, Fernando Mandoki, Juan José |
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Keywords | Flow cytometry Cytostatic effect G 1/S transition Cyclin D1 7-Hydroxycoumarin Cell cycle Antineoplastic agent Human Cell proliferation Lung Cyclin Activity Product Malignant tumor Information Coumarin In vitro Mechanism In vivo Type Cell line Decrease Inhibition Tumor cell Gain |
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SubjectTerms | 7-Hydroxycoumarin Adenocarcinoma - pathology Antineoplastic Agents - pharmacology Biological and medical sciences Biotransformation Cell cycle Cell Cycle - drug effects Cell Division Coumarins - pharmacology Cyclin D1 Cyclin D1 - biosynthesis Cytostatic effect Dose-Response Relationship, Drug Flow cytometry G 1/S transition Gene Expression Regulation, Neoplastic Humans Lung Neoplasms - pathology Medical sciences Pneumology Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger - biosynthesis Tumor Cells, Cultured Tumors of the respiratory system and mediastinum Umbelliferones - pharmacology |
Title | Decrease of cyclin D1 in the human lung adenocarcinoma cell line A-427 by 7-hydroxycoumarin |
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