Expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cells
Statins have anticancer effects and may be used as anticancer agents via drug repositioning. In reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays, the internal reference gene must not be affected by any experimental conditions. As statins exert a wide range of effects on...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-07-01
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| Series: | Heliyon |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405844023052258 |
| _version_ | 1797771444359266304 |
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| author | Nanami Irie Katsuhiko Warita Jiro Tashiro Yaxuan Zhou Takuro Ishikawa Zoltán N. Oltvai Tomoko Warita |
| author_facet | Nanami Irie Katsuhiko Warita Jiro Tashiro Yaxuan Zhou Takuro Ishikawa Zoltán N. Oltvai Tomoko Warita |
| author_sort | Nanami Irie |
| collection | DOAJ |
| description | Statins have anticancer effects and may be used as anticancer agents via drug repositioning. In reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays, the internal reference gene must not be affected by any experimental conditions. As statins exert a wide range of effects on cells by inhibiting the mevalonate pathway, it is possible that statin treatment might alter the expression of housekeeping genes used as internal reference genes, thereby misleading the assessment of obtained gene expression data. Here, we evaluated the expression stability of internal reference genes in atorvastatin-treated cancer cell lines. We treated both statin-sensitive and statin-resistant cancer cell lines with atorvastatin at seven different concentrations and performed RT-qPCR on 15 housekeeping genes whose expression stability was then assessed using five different algorithms. In both statin-sensitive and statin-resistant cancer cell lines, atorvastatin affected the expression of certain internal reference genes in a dose-dependent and cancer cell line-dependent manner; therefore, caution should be exercised when comparing target gene expression between cells. Our findings emphasize the importance of the validation of internal reference genes in gene expression analyses in drug treatment-based cancer research. |
| first_indexed | 2024-03-12T21:37:40Z |
| format | Article |
| id | doaj.art-9c45f93c82474f968d12c4185f8f9a29 |
| institution | Directory Open Access Journal |
| issn | 2405-8440 |
| language | English |
| last_indexed | 2024-03-12T21:37:40Z |
| publishDate | 2023-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Heliyon |
| spelling | doaj.art-9c45f93c82474f968d12c4185f8f9a292023-07-27T05:58:16ZengElsevierHeliyon2405-84402023-07-0197e18017Expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cellsNanami Irie0Katsuhiko Warita1Jiro Tashiro2Yaxuan Zhou3Takuro Ishikawa4Zoltán N. Oltvai5Tomoko Warita6Graduate School of Science and Technology, Kwansei Gakuin University, 1 Gakuen Uegahara, Sanda, Hyogo 669‐1330, JapanDepartment of Veterinary Anatomy, School of Veterinary Medicine, Tottori University, 4-101 Koyama Minami, Tottori, Tottori 680-8553, Japan; Corresponding author.Department of Veterinary Anatomy, School of Veterinary Medicine, Tottori University, 4-101 Koyama Minami, Tottori, Tottori 680-8553, JapanGraduate School of Science and Technology, Kwansei Gakuin University, 1 Gakuen Uegahara, Sanda, Hyogo 669‐1330, JapanDepartment of Anatomy, School of Medicine, Aichi Medical University, 1-1 Yazakokarimata, Nagakute, Aichi 480-1195, JapanDepartment of Pathology and Laboratory Medicine, University of Rochester, 601 Elmwood Ave., Rochester, NY 14642, USADepartment of Biomedical Sciences, School of Biological and Environmental Sciences, Kwansei Gakuin University, 1 Gakuen Uegahara, Sanda, Hyogo 669‐1330, JapanStatins have anticancer effects and may be used as anticancer agents via drug repositioning. In reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays, the internal reference gene must not be affected by any experimental conditions. As statins exert a wide range of effects on cells by inhibiting the mevalonate pathway, it is possible that statin treatment might alter the expression of housekeeping genes used as internal reference genes, thereby misleading the assessment of obtained gene expression data. Here, we evaluated the expression stability of internal reference genes in atorvastatin-treated cancer cell lines. We treated both statin-sensitive and statin-resistant cancer cell lines with atorvastatin at seven different concentrations and performed RT-qPCR on 15 housekeeping genes whose expression stability was then assessed using five different algorithms. In both statin-sensitive and statin-resistant cancer cell lines, atorvastatin affected the expression of certain internal reference genes in a dose-dependent and cancer cell line-dependent manner; therefore, caution should be exercised when comparing target gene expression between cells. Our findings emphasize the importance of the validation of internal reference genes in gene expression analyses in drug treatment-based cancer research.http://www.sciencedirect.com/science/article/pii/S2405844023052258Mevalonate pathwayCancer cellsStatinsHousekeeping genesRT-qPCROptimal internal reference genes |
| spellingShingle | Nanami Irie Katsuhiko Warita Jiro Tashiro Yaxuan Zhou Takuro Ishikawa Zoltán N. Oltvai Tomoko Warita Expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cells Heliyon Mevalonate pathway Cancer cells Statins Housekeeping genes RT-qPCR Optimal internal reference genes |
| title | Expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cells |
| title_full | Expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cells |
| title_fullStr | Expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cells |
| title_full_unstemmed | Expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cells |
| title_short | Expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cells |
| title_sort | expression of housekeeping genes varies depending on mevalonate pathway inhibition in cancer cells |
| topic | Mevalonate pathway Cancer cells Statins Housekeeping genes RT-qPCR Optimal internal reference genes |
| url | http://www.sciencedirect.com/science/article/pii/S2405844023052258 |
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