Deficiency of RARα Suppresses Decidualization via Downregulating CEBPB Transcription in Women With Recurrent Implantation Failure
BackgroundRecurrent implantation failure (RIF) is a disease associated with endometrial receptivity dysfunction. Retinoic acid receptor alpha (RARα) is an important protein in many biological processes, such as differentiation and development. However, the exact underlying mechanism whereby RARα aff...
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Endocrinology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fendo.2022.753416/full |
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| author | Caiyi Huang Caiyi Huang Caiyi Huang Caiyi Huang Caiyi Huang Qian Zhang Qian Zhang Qian Zhang Tianxiang Ni Tianxiang Ni Tianxiang Ni Tingting Zhou Tingting Zhou Tingting Zhou Chunzi Lv Chunzi Lv Chunzi Lv Yan Li Yan Li Yan Li Junhao Yan Junhao Yan Junhao Yan Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen |
| author_facet | Caiyi Huang Caiyi Huang Caiyi Huang Caiyi Huang Caiyi Huang Qian Zhang Qian Zhang Qian Zhang Tianxiang Ni Tianxiang Ni Tianxiang Ni Tingting Zhou Tingting Zhou Tingting Zhou Chunzi Lv Chunzi Lv Chunzi Lv Yan Li Yan Li Yan Li Junhao Yan Junhao Yan Junhao Yan Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen |
| author_sort | Caiyi Huang |
| collection | DOAJ |
| description | BackgroundRecurrent implantation failure (RIF) is a disease associated with endometrial receptivity dysfunction. Retinoic acid receptor alpha (RARα) is an important protein in many biological processes, such as differentiation and development. However, the exact underlying mechanism whereby RARα affects RIF remains unknown. This study investigated RARα expression and its contribution in the mid-luteal phase endometria of patients with RIF.MethodsThe expression levels of RARα and CCAAT/enhancer-binding protein (C/EBP) β in the endometria of the RIF and normal group were investigated using western blotting and immunohistochemistry. In in vitro experiments, immortal telomerase-transformed human endometrial stromal cells (T-HESCs) were incubated with medroxyprogesterone-17-acetate (MPA) and cyclic adenosine monophosphate (cAMP) for 4 days to induce decidualization. The expression levels of the decidualization markers prolactin (PRL) and insulin-like growth factor-binding protein-1 (IGFBP-1) were determined using quantitative polymerase chain reaction. RARα was knocked down using a small interfering RNA, and C/EBPβ was overexpressed from an adenoviral vector. The transcriptional regulation of CEBPB by RARα was determined by chromatin immunoprecipitation (ChIP) assay and luciferase assays.ResultsWe found that the expression levels of RARα decreased in the mid-luteal endometria of RIF patients. After 4 days of decidualization induction in vitro, RARα knockdown impaired the decidualization of T-HESCs and downregulated the expression of C/EBPβ. The restoration of C/EBPβ expression rescued the RARα knockdown-induced suppression of T-HESC decidualization. In ChIP analysis of lysates from decidualized T-HESCs, the CEBPB promoter region was enriched in chromatin fragments pulled down using an anti-RARα antibody. However, the relationship between CEBPB transcription and RARα expression levels was only observed when the decidualization of T-HESCs was induced by the addition of cAMP and MPA. To identify the binding site of RARα/retinoid X receptor α, we performed luciferase assays. Mutation of the predicted binding site in CEBPB (-2,009/-1,781) decreased the transcriptional activity of the reporter. To confirm this mechanism, the expression levels of C/EBPβ in the mid-luteal endometria of RIF patients were determined and found to decrease with decreased RARα expression levels.ConclusionA deficiency of RARα expression in the mid-luteal endometrium inhibits decidualization due to the downregulation of CEBPB transcription. This is a potential mechanism contributing to RIF. |
| first_indexed | 2024-04-14T00:22:46Z |
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| issn | 1664-2392 |
| language | English |
| last_indexed | 2024-04-14T00:22:46Z |
| publishDate | 2022-05-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Endocrinology |
| spelling | doaj.art-5a63d7d085d1452d85269af1b60904dc2022-12-22T02:22:54ZengFrontiers Media S.A.Frontiers in Endocrinology1664-23922022-05-011310.3389/fendo.2022.753416753416Deficiency of RARα Suppresses Decidualization via Downregulating CEBPB Transcription in Women With Recurrent Implantation FailureCaiyi Huang0Caiyi Huang1Caiyi Huang2Caiyi Huang3Caiyi Huang4Qian Zhang5Qian Zhang6Qian Zhang7Tianxiang Ni8Tianxiang Ni9Tianxiang Ni10Tingting Zhou11Tingting Zhou12Tingting Zhou13Chunzi Lv14Chunzi Lv15Chunzi Lv16Yan Li17Yan Li18Yan Li19Junhao Yan20Junhao Yan21Junhao Yan22Zi-Jiang Chen23Zi-Jiang Chen24Zi-Jiang Chen25Zi-Jiang Chen26Zi-Jiang Chen27Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai Jiao Tong University, Shanghai, ChinaCenter for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, ChinaNational Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, ChinaKey Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, ChinaCenter for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, ChinaNational Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, ChinaKey Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, ChinaCenter for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, ChinaNational Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, ChinaKey Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, ChinaCenter for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, ChinaNational Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, ChinaKey Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, ChinaCenter for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, ChinaNational Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, ChinaKey Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, ChinaCenter for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, ChinaNational Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, ChinaKey Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, ChinaCenter for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, ChinaNational Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, ChinaKey Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, ChinaCenter for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, ChinaShanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai Jiao Tong University, Shanghai, ChinaCenter for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, ChinaNational Research Center for Assisted Reproductive Technology and Reproductive Genetics, Shandong University, Jinan, ChinaKey Laboratory of Reproductive Endocrinology of Ministry of Education, Shandong University, Jinan, ChinaBackgroundRecurrent implantation failure (RIF) is a disease associated with endometrial receptivity dysfunction. Retinoic acid receptor alpha (RARα) is an important protein in many biological processes, such as differentiation and development. However, the exact underlying mechanism whereby RARα affects RIF remains unknown. This study investigated RARα expression and its contribution in the mid-luteal phase endometria of patients with RIF.MethodsThe expression levels of RARα and CCAAT/enhancer-binding protein (C/EBP) β in the endometria of the RIF and normal group were investigated using western blotting and immunohistochemistry. In in vitro experiments, immortal telomerase-transformed human endometrial stromal cells (T-HESCs) were incubated with medroxyprogesterone-17-acetate (MPA) and cyclic adenosine monophosphate (cAMP) for 4 days to induce decidualization. The expression levels of the decidualization markers prolactin (PRL) and insulin-like growth factor-binding protein-1 (IGFBP-1) were determined using quantitative polymerase chain reaction. RARα was knocked down using a small interfering RNA, and C/EBPβ was overexpressed from an adenoviral vector. The transcriptional regulation of CEBPB by RARα was determined by chromatin immunoprecipitation (ChIP) assay and luciferase assays.ResultsWe found that the expression levels of RARα decreased in the mid-luteal endometria of RIF patients. After 4 days of decidualization induction in vitro, RARα knockdown impaired the decidualization of T-HESCs and downregulated the expression of C/EBPβ. The restoration of C/EBPβ expression rescued the RARα knockdown-induced suppression of T-HESC decidualization. In ChIP analysis of lysates from decidualized T-HESCs, the CEBPB promoter region was enriched in chromatin fragments pulled down using an anti-RARα antibody. However, the relationship between CEBPB transcription and RARα expression levels was only observed when the decidualization of T-HESCs was induced by the addition of cAMP and MPA. To identify the binding site of RARα/retinoid X receptor α, we performed luciferase assays. Mutation of the predicted binding site in CEBPB (-2,009/-1,781) decreased the transcriptional activity of the reporter. To confirm this mechanism, the expression levels of C/EBPβ in the mid-luteal endometria of RIF patients were determined and found to decrease with decreased RARα expression levels.ConclusionA deficiency of RARα expression in the mid-luteal endometrium inhibits decidualization due to the downregulation of CEBPB transcription. This is a potential mechanism contributing to RIF.https://www.frontiersin.org/articles/10.3389/fendo.2022.753416/fullRARαrecurrent implantation failure (RIF)decidualizationCEBPBendometria stromal cells |
| spellingShingle | Caiyi Huang Caiyi Huang Caiyi Huang Caiyi Huang Caiyi Huang Qian Zhang Qian Zhang Qian Zhang Tianxiang Ni Tianxiang Ni Tianxiang Ni Tingting Zhou Tingting Zhou Tingting Zhou Chunzi Lv Chunzi Lv Chunzi Lv Yan Li Yan Li Yan Li Junhao Yan Junhao Yan Junhao Yan Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen Zi-Jiang Chen Deficiency of RARα Suppresses Decidualization via Downregulating CEBPB Transcription in Women With Recurrent Implantation Failure Frontiers in Endocrinology RARα recurrent implantation failure (RIF) decidualization CEBPB endometria stromal cells |
| title | Deficiency of RARα Suppresses Decidualization via Downregulating CEBPB Transcription in Women With Recurrent Implantation Failure |
| title_full | Deficiency of RARα Suppresses Decidualization via Downregulating CEBPB Transcription in Women With Recurrent Implantation Failure |
| title_fullStr | Deficiency of RARα Suppresses Decidualization via Downregulating CEBPB Transcription in Women With Recurrent Implantation Failure |
| title_full_unstemmed | Deficiency of RARα Suppresses Decidualization via Downregulating CEBPB Transcription in Women With Recurrent Implantation Failure |
| title_short | Deficiency of RARα Suppresses Decidualization via Downregulating CEBPB Transcription in Women With Recurrent Implantation Failure |
| title_sort | deficiency of rarα suppresses decidualization via downregulating cebpb transcription in women with recurrent implantation failure |
| topic | RARα recurrent implantation failure (RIF) decidualization CEBPB endometria stromal cells |
| url | https://www.frontiersin.org/articles/10.3389/fendo.2022.753416/full |
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