The effect of AP-2δ on transcription of the Prestin gene in HEI-OC1 cells upon oxidative stress
Abstract Background The study aimed to investigate the effect of oxidative stress on Prestin expression, and explore the transcription factors (TFs) that are involved in regulating the expression of Prestin in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells upon oxidative stress. Methods Quanti...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-06-01
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| Series: | Cellular & Molecular Biology Letters |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s11658-019-0170-0 |
| _version_ | 1818965042330075136 |
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| author | Xuan Luo Yun Xia Xu-Dong Li Jun-Yi Wang |
| author_facet | Xuan Luo Yun Xia Xu-Dong Li Jun-Yi Wang |
| author_sort | Xuan Luo |
| collection | DOAJ |
| description | Abstract Background The study aimed to investigate the effect of oxidative stress on Prestin expression, and explore the transcription factors (TFs) that are involved in regulating the expression of Prestin in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells upon oxidative stress. Methods Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression level of Prestin. Reverse chromatin immunoprecipitation (reverse ChIP) assay was performed to identify proteins that could bind to the Prestin gene. Small interfering RNA (siRNA) and chromatin immunoprecipitation (ChIP) experiments were used to further verify the results. HEI-OC1 cells were incubated with four different concentrations of tert-butyl hydroperoxide (t-BHP) for 24 h or 48 h to construct the oxidative stress model. Results Oxidative stress induced Prestin increase at the mRNA level but with a concomitant decrease at the protein level. TF activating enhancer binding protein-2δ (AP-2δ) screened by reverse ChIP assay was demonstrated to bind to transcriptional start site 1441 of the Prestin promoter region and negatively regulate the expression of Prestin by siRNA and ChIP experiments. Furthermore, AP-2δ was down-regulated under oxidative stress. Conclusions In conclusion, oxidative stress inhibits the expression of Prestin protein, and the transcription mechanism is triggered to compensate for the loss of Prestin protein. AP-2δ is one of the important TFs that suppresses transcription of the Prestin gene, and AP-2δ suppression further boosted Prestin mRNA activation under oxidative stress. |
| first_indexed | 2024-12-20T13:10:43Z |
| format | Article |
| id | doaj.art-4baa81a691c64a8d83613ee00377ec35 |
| institution | Directory Open Access Journal |
| issn | 1425-8153 1689-1392 |
| language | English |
| last_indexed | 2024-12-20T13:10:43Z |
| publishDate | 2019-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Cellular & Molecular Biology Letters |
| spelling | doaj.art-4baa81a691c64a8d83613ee00377ec352022-12-21T19:39:40ZengBMCCellular & Molecular Biology Letters1425-81531689-13922019-06-0124111410.1186/s11658-019-0170-0The effect of AP-2δ on transcription of the Prestin gene in HEI-OC1 cells upon oxidative stressXuan Luo0Yun Xia1Xu-Dong Li2Jun-Yi Wang3Department of Labor Health and Environmental Hygiene, School of Public Health, Guangdong Pharmaceutical UniversityDepartment of Labor Health and Environmental Hygiene, School of Public Health, Guangdong Pharmaceutical UniversityKey Laboratory, Occupational Disease Prevention and Control of Hospital of Guangdong ProvinceDepartment of Labor Health and Environmental Hygiene, School of Public Health, Guangdong Pharmaceutical UniversityAbstract Background The study aimed to investigate the effect of oxidative stress on Prestin expression, and explore the transcription factors (TFs) that are involved in regulating the expression of Prestin in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells upon oxidative stress. Methods Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression level of Prestin. Reverse chromatin immunoprecipitation (reverse ChIP) assay was performed to identify proteins that could bind to the Prestin gene. Small interfering RNA (siRNA) and chromatin immunoprecipitation (ChIP) experiments were used to further verify the results. HEI-OC1 cells were incubated with four different concentrations of tert-butyl hydroperoxide (t-BHP) for 24 h or 48 h to construct the oxidative stress model. Results Oxidative stress induced Prestin increase at the mRNA level but with a concomitant decrease at the protein level. TF activating enhancer binding protein-2δ (AP-2δ) screened by reverse ChIP assay was demonstrated to bind to transcriptional start site 1441 of the Prestin promoter region and negatively regulate the expression of Prestin by siRNA and ChIP experiments. Furthermore, AP-2δ was down-regulated under oxidative stress. Conclusions In conclusion, oxidative stress inhibits the expression of Prestin protein, and the transcription mechanism is triggered to compensate for the loss of Prestin protein. AP-2δ is one of the important TFs that suppresses transcription of the Prestin gene, and AP-2δ suppression further boosted Prestin mRNA activation under oxidative stress.http://link.springer.com/article/10.1186/s11658-019-0170-0PrestinAP-2δOxidative stressTranscriptional regulationHEI-OC1 cells |
| spellingShingle | Xuan Luo Yun Xia Xu-Dong Li Jun-Yi Wang The effect of AP-2δ on transcription of the Prestin gene in HEI-OC1 cells upon oxidative stress Cellular & Molecular Biology Letters Prestin AP-2δ Oxidative stress Transcriptional regulation HEI-OC1 cells |
| title | The effect of AP-2δ on transcription of the Prestin gene in HEI-OC1 cells upon oxidative stress |
| title_full | The effect of AP-2δ on transcription of the Prestin gene in HEI-OC1 cells upon oxidative stress |
| title_fullStr | The effect of AP-2δ on transcription of the Prestin gene in HEI-OC1 cells upon oxidative stress |
| title_full_unstemmed | The effect of AP-2δ on transcription of the Prestin gene in HEI-OC1 cells upon oxidative stress |
| title_short | The effect of AP-2δ on transcription of the Prestin gene in HEI-OC1 cells upon oxidative stress |
| title_sort | effect of ap 2δ on transcription of the prestin gene in hei oc1 cells upon oxidative stress |
| topic | Prestin AP-2δ Oxidative stress Transcriptional regulation HEI-OC1 cells |
| url | http://link.springer.com/article/10.1186/s11658-019-0170-0 |
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