Induction of Continuous Expression of Cyclooxygenase -2 in Human Breast Cancer Cell Line Using COX-2 cDNA Plasmid Transfection
Background and purpose: One of the major problems in chemotherapy is resistance to anti-cancer drugs. Recent studies have demonstrated a relationship between cyclooxygenase-2 (COX-2) expression and progression of multidrug resistance (MDR). One of the proteins involved in drug resistance is ABCG2 (A...
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| Format: | Article |
| Language: | English |
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Mazandaran University of Medical Sciences
2017-11-01
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| Series: | Journal of Mazandaran University of Medical Sciences |
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| Online Access: | http://jmums.mazums.ac.ir/article-1-6605-en.html |
| _version_ | 1797943744487489536 |
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| author | Saeedeh Vahedi BiBi Fatemeh Kalalinia Zahra Noormohamadi Fatemeh Mosaffa Javad Behravan |
| author_facet | Saeedeh Vahedi BiBi Fatemeh Kalalinia Zahra Noormohamadi Fatemeh Mosaffa Javad Behravan |
| author_sort | Saeedeh Vahedi |
| collection | DOAJ |
| description | Background and purpose: One of the major problems in chemotherapy is resistance to anti-cancer drugs. Recent studies have demonstrated a relationship between cyclooxygenase-2 (COX-2) expression and progression of multidrug resistance (MDR). One of the proteins involved in drug resistance is ABCG2 (ATP-binding cassette sub-family G member 2) that is often overexpressed in patients with cancer. This study was performed to develop the MCF7 and MCF7/MX human breast cancer cell line which continuously expressed COX-2 in order to evaluate the effect of induction of COX-2 expression on ABCG2 expression.
Materials and methods: For this purpose, a plasmid containing the sequence of cyclooxygenase-2 was cloned in E. coli. The purified plasmid was then transferred into the MCF7 and MCF7/MX cell lines by transfection process. Finally, the protein content of transfected cells were extracted and analyzed by Western blotting. The COX-2 expression was compared between transfected cells.
Results: The results showed that COX-2 protein levels were the highest in MCF7 cells transfected with a DNA and Fusion 6 ratio of 3:1 and the medium containing genticine added after 24 hours. Also, compared to the control cell line, in MCF7/MX cells the expression of COX-2 gene increased in transfected cells with a DNA and Fusion 6 ratio of 3:2 (P<0.05).
Conclusion: In fact, the level of target gene expression may vary based on the cell line and culture conditions. Increase in the amount of DNA does not necessarily lead to increase in transfection efficiency. |
| first_indexed | 2024-04-10T20:29:02Z |
| format | Article |
| id | doaj.art-e25debd7bd944dc8b24fe065c26932e9 |
| institution | Directory Open Access Journal |
| issn | 1735-9260 1735-9279 |
| language | English |
| last_indexed | 2024-04-10T20:29:02Z |
| publishDate | 2017-11-01 |
| publisher | Mazandaran University of Medical Sciences |
| record_format | Article |
| series | Journal of Mazandaran University of Medical Sciences |
| spelling | doaj.art-e25debd7bd944dc8b24fe065c26932e92023-01-25T07:50:07ZengMazandaran University of Medical SciencesJournal of Mazandaran University of Medical Sciences1735-92601735-92792017-11-01271543850Induction of Continuous Expression of Cyclooxygenase -2 in Human Breast Cancer Cell Line Using COX-2 cDNA Plasmid TransfectionSaeedeh Vahedi0BiBi Fatemeh Kalalinia1Zahra Noormohamadi2Fatemeh Mosaffa3Javad Behravan4 MSc in Genetics, Faculty of Sciences, Islamic Azad University, Science and Research Branch, Tehran, Iran Assistant Professor, Biotechnology Research Center, Faculty of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran Associate Professor, Department of Biology, Faculty of Sciences, Islamic Azad University, Science and Research Branch, Tehran, Iran Associate Professor, Biotechnology Research Center, Faculty of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran Professor, Biotechnology Research Center, Faculty of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran Background and purpose: One of the major problems in chemotherapy is resistance to anti-cancer drugs. Recent studies have demonstrated a relationship between cyclooxygenase-2 (COX-2) expression and progression of multidrug resistance (MDR). One of the proteins involved in drug resistance is ABCG2 (ATP-binding cassette sub-family G member 2) that is often overexpressed in patients with cancer. This study was performed to develop the MCF7 and MCF7/MX human breast cancer cell line which continuously expressed COX-2 in order to evaluate the effect of induction of COX-2 expression on ABCG2 expression. Materials and methods: For this purpose, a plasmid containing the sequence of cyclooxygenase-2 was cloned in E. coli. The purified plasmid was then transferred into the MCF7 and MCF7/MX cell lines by transfection process. Finally, the protein content of transfected cells were extracted and analyzed by Western blotting. The COX-2 expression was compared between transfected cells. Results: The results showed that COX-2 protein levels were the highest in MCF7 cells transfected with a DNA and Fusion 6 ratio of 3:1 and the medium containing genticine added after 24 hours. Also, compared to the control cell line, in MCF7/MX cells the expression of COX-2 gene increased in transfected cells with a DNA and Fusion 6 ratio of 3:2 (P<0.05). Conclusion: In fact, the level of target gene expression may vary based on the cell line and culture conditions. Increase in the amount of DNA does not necessarily lead to increase in transfection efficiency.http://jmums.mazums.ac.ir/article-1-6605-en.htmlmultidrug resistancecox-2mcf7 cell line |
| spellingShingle | Saeedeh Vahedi BiBi Fatemeh Kalalinia Zahra Noormohamadi Fatemeh Mosaffa Javad Behravan Induction of Continuous Expression of Cyclooxygenase -2 in Human Breast Cancer Cell Line Using COX-2 cDNA Plasmid Transfection Journal of Mazandaran University of Medical Sciences multidrug resistance cox-2 mcf7 cell line |
| title | Induction of Continuous Expression of Cyclooxygenase -2 in Human Breast Cancer Cell Line Using COX-2 cDNA Plasmid Transfection |
| title_full | Induction of Continuous Expression of Cyclooxygenase -2 in Human Breast Cancer Cell Line Using COX-2 cDNA Plasmid Transfection |
| title_fullStr | Induction of Continuous Expression of Cyclooxygenase -2 in Human Breast Cancer Cell Line Using COX-2 cDNA Plasmid Transfection |
| title_full_unstemmed | Induction of Continuous Expression of Cyclooxygenase -2 in Human Breast Cancer Cell Line Using COX-2 cDNA Plasmid Transfection |
| title_short | Induction of Continuous Expression of Cyclooxygenase -2 in Human Breast Cancer Cell Line Using COX-2 cDNA Plasmid Transfection |
| title_sort | induction of continuous expression of cyclooxygenase 2 in human breast cancer cell line using cox 2 cdna plasmid transfection |
| topic | multidrug resistance cox-2 mcf7 cell line |
| url | http://jmums.mazums.ac.ir/article-1-6605-en.html |
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