Irreversible methadone-induced GSTP1 downregulation in SH-SY5Y cells
Abstract Background Methadone has been reported to downregulate the expression of glutathione S-transferase P1 (GSTP1) among nine antioxidant genes in SH-SY5Y cells after both short- and long-term treatment. GSTP1 plays a key role in the detoxification of many xenobiotics and is frequently associate...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2024-03-01
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| Series: | Egyptian Journal of Medical Human Genetics |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s43042-024-00504-7 |
| _version_ | 1797259226513408000 |
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| author | Khyber Saify Mostafa Saadat |
| author_facet | Khyber Saify Mostafa Saadat |
| author_sort | Khyber Saify |
| collection | DOAJ |
| description | Abstract Background Methadone has been reported to downregulate the expression of glutathione S-transferase P1 (GSTP1) among nine antioxidant genes in SH-SY5Y cells after both short- and long-term treatment. GSTP1 plays a key role in the detoxification of many xenobiotics and is frequently associated with various diseases, especially tumors. The objective of this study is to determine whether this change is reversible. Methods Two different treatment protocols were used. The first protocol evaluated the reversibility of the GSTP1 mRNA change, while the second protocol evaluated the methylation status of the GSTP1 promoter site. To investigate the reversibility of the GSTP1 mRNA change, SH-SY5Y cells were treated with methadone. The drug was then removed from the medium and the cells were cultured in methadone-free medium for a period of time. GSTP1 mRNA levels were expressed as cycle threshold (Ct) values using TATA box-binding protein as a calibrator gene. Methylation at the promoter site was detected by bisulfite treatment. Results The analysis of variance revealed no significant change in GSTP1 mRNA levels in the cells after methadone was removed from the medium of methadone-treated cells. The study also examined the methylation status of a CpG island in the promoter of GSTP1 in the treated cells. The results demonstrate that although methadone downregulates the mRNA level of GSTP1 in treated cells, it does not induce methylation in the GSTP1 promoter region. Conclusions The expression of the GSTP1 remains downregulated even after methadone removal from SH-SY5Y cell culture medium; however, methylation of the GSTP1 promoter site does not play a role in this process. |
| first_indexed | 2024-04-24T23:06:04Z |
| format | Article |
| id | doaj.art-8b55ffeee5a14d03aa0f479bc39f290c |
| institution | Directory Open Access Journal |
| issn | 2090-2441 |
| language | English |
| last_indexed | 2024-04-24T23:06:04Z |
| publishDate | 2024-03-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Egyptian Journal of Medical Human Genetics |
| spelling | doaj.art-8b55ffeee5a14d03aa0f479bc39f290c2024-03-17T12:28:11ZengSpringerOpenEgyptian Journal of Medical Human Genetics2090-24412024-03-012511710.1186/s43042-024-00504-7Irreversible methadone-induced GSTP1 downregulation in SH-SY5Y cellsKhyber Saify0Mostafa Saadat1Department of Biology, School of Science, Shiraz UniversityDepartment of Biology, School of Science, Shiraz UniversityAbstract Background Methadone has been reported to downregulate the expression of glutathione S-transferase P1 (GSTP1) among nine antioxidant genes in SH-SY5Y cells after both short- and long-term treatment. GSTP1 plays a key role in the detoxification of many xenobiotics and is frequently associated with various diseases, especially tumors. The objective of this study is to determine whether this change is reversible. Methods Two different treatment protocols were used. The first protocol evaluated the reversibility of the GSTP1 mRNA change, while the second protocol evaluated the methylation status of the GSTP1 promoter site. To investigate the reversibility of the GSTP1 mRNA change, SH-SY5Y cells were treated with methadone. The drug was then removed from the medium and the cells were cultured in methadone-free medium for a period of time. GSTP1 mRNA levels were expressed as cycle threshold (Ct) values using TATA box-binding protein as a calibrator gene. Methylation at the promoter site was detected by bisulfite treatment. Results The analysis of variance revealed no significant change in GSTP1 mRNA levels in the cells after methadone was removed from the medium of methadone-treated cells. The study also examined the methylation status of a CpG island in the promoter of GSTP1 in the treated cells. The results demonstrate that although methadone downregulates the mRNA level of GSTP1 in treated cells, it does not induce methylation in the GSTP1 promoter region. Conclusions The expression of the GSTP1 remains downregulated even after methadone removal from SH-SY5Y cell culture medium; however, methylation of the GSTP1 promoter site does not play a role in this process.https://doi.org/10.1186/s43042-024-00504-7GSTP1MethylationMethadoneSH-SY5Y |
| spellingShingle | Khyber Saify Mostafa Saadat Irreversible methadone-induced GSTP1 downregulation in SH-SY5Y cells Egyptian Journal of Medical Human Genetics GSTP1 Methylation Methadone SH-SY5Y |
| title | Irreversible methadone-induced GSTP1 downregulation in SH-SY5Y cells |
| title_full | Irreversible methadone-induced GSTP1 downregulation in SH-SY5Y cells |
| title_fullStr | Irreversible methadone-induced GSTP1 downregulation in SH-SY5Y cells |
| title_full_unstemmed | Irreversible methadone-induced GSTP1 downregulation in SH-SY5Y cells |
| title_short | Irreversible methadone-induced GSTP1 downregulation in SH-SY5Y cells |
| title_sort | irreversible methadone induced gstp1 downregulation in sh sy5y cells |
| topic | GSTP1 Methylation Methadone SH-SY5Y |
| url | https://doi.org/10.1186/s43042-024-00504-7 |
| work_keys_str_mv | AT khybersaify irreversiblemethadoneinducedgstp1downregulationinshsy5ycells AT mostafasaadat irreversiblemethadoneinducedgstp1downregulationinshsy5ycells |