CpG site hypomethylation at ETS1‑binding region regulates DLK1 expression in Chinese patients with Tetralogy of Fallot
Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart malformation accounting for ~10% of cases. Although the pathogenesis of TOF is complex and largely unknown, epigenetics plays a huge role, specifically DNA methylation. The protein δ like non‑canonical Notch ligand 1 (DLK1) gene...
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| Veröffentlicht in: | Molecular medicine reports 2022-03, Vol.25 (3), Article 93 |
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| creator | Tian, Guixiang He, Lili Gu, Ruoyi Sun, Jingwei Chen, Weicheng Qian, Yanyan Ma, Xiaojing Yan, Weili Zhao, Zhenshan Xu, Ziqing Suo, Meijiao Sheng, Wei Huang, Guoying |
| description | Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart malformation accounting for ~10% of cases. Although the pathogenesis of TOF is complex and largely unknown, epigenetics plays a huge role, specifically DNA methylation. The protein δ like non‑canonical Notch ligand 1 (DLK1) gene encodes a non‑canonical ligand of the Notch signaling pathway, which is involved in heart development. However, the epigenetic mechanism of DLK1 in the pathogenesis of TOF is yet to be elucidated. Therefore, the present study aimed to clarify its specific mechanism. In this study, immunohistochemistry was used to detect the protein expression of DLK1 and the methylation status of the DLK1 promoter was measured via bisulfite sequencing PCR. Dual‑luciferase reporter assays were performed to examine the influence of transcription factor ETS proto‑oncogene 1 (ETS1) on DLK1 gene expression. The electrophoretic mobility shift assay and chromatin immunoprecipitation assay, both
and
, were used to verify the binding of the ETS1 transcription factor to the DLK1 promoter as well as the influence of methylation status of DLK1 promoter on this binding affinity. The expression of DLK1 in the right ventricular outflow tract was significantly lower in patients with Tetralogy of Fallot (TOF) than that in controls (P |
| doi_str_mv | 10.3892/mmr.2022.12609 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_8809049</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A695364047</galeid><sourcerecordid>A695364047</sourcerecordid><originalsourceid>FETCH-LOGICAL-c485t-5b03cddd9716c26ea106222128c6a161a5884cf3a474ef8b0009f7fdebe73cb93</originalsourceid><addsrcrecordid>eNptkk1v1DAQhiMEoqVw5YgsceGyi7_i2BekamkLYiUOLGfLcSZZV4kdYge6N_4Cf5FfggNL-VDlw1ieZ96Zsd6ieErwmklFXw7DtKaY0jWhAqt7xSmpFFkxjPn9450qVZ0Uj2K8xliUtFQPixNW4lJVnJ8WN5vxCkWXAO0PYxgg7Q-9SS54ZBK62H0g379-q51vnO_QBN2SyGHODET0evuOILgZJ4hxyTiPNnvnIQIaswj4FNEXl_ZoB2kyfegOKLTo0vR9SI-LB63pIzw5xrPi4-XFbvNmtX1_9XZzvl1ZLsu0KmvMbNM0qiLCUgGGYEEpJVRaYYggppSS25YZXnFoZY0xVm3VNlBDxWyt2Fnx6pfuONcDNDYPlUfR4-QGMx10ME7_m_Fur7vwWUuJFeaLwIujwBQ-zRCTHly00PfGQ5ijpss8kpGSZfT5f-h1mCef11uoivFSSPGH6kwP2vk25L52EdXnQpVMcMyrTK3voPJpYHA2eGhdfr-rwE4hxgna2x0J1otXdPaKXryif3olFzz7-2du8d_mYD8Aq4W7lQ</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2627345686</pqid></control><display><type>article</type><title>CpG site hypomethylation at ETS1‑binding region regulates DLK1 expression in Chinese patients with Tetralogy of Fallot</title><source>Spandidos Publications Journals</source><source>MEDLINE</source><source>Alma/SFX Local Collection</source><source>EZB Electronic Journals Library</source><creator>Tian, Guixiang ; He, Lili ; Gu, Ruoyi ; Sun, Jingwei ; Chen, Weicheng ; Qian, Yanyan ; Ma, Xiaojing ; Yan, Weili ; Zhao, Zhenshan ; Xu, Ziqing ; Suo, Meijiao ; Sheng, Wei ; Huang, Guoying</creator><creatorcontrib>Tian, Guixiang ; He, Lili ; Gu, Ruoyi ; Sun, Jingwei ; Chen, Weicheng ; Qian, Yanyan ; Ma, Xiaojing ; Yan, Weili ; Zhao, Zhenshan ; Xu, Ziqing ; Suo, Meijiao ; Sheng, Wei ; Huang, Guoying</creatorcontrib><description>Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart malformation accounting for ~10% of cases. Although the pathogenesis of TOF is complex and largely unknown, epigenetics plays a huge role, specifically DNA methylation. The protein δ like non‑canonical Notch ligand 1 (DLK1) gene encodes a non‑canonical ligand of the Notch signaling pathway, which is involved in heart development. However, the epigenetic mechanism of DLK1 in the pathogenesis of TOF is yet to be elucidated. Therefore, the present study aimed to clarify its specific mechanism. In this study, immunohistochemistry was used to detect the protein expression of DLK1 and the methylation status of the DLK1 promoter was measured via bisulfite sequencing PCR. Dual‑luciferase reporter assays were performed to examine the influence of transcription factor ETS proto‑oncogene 1 (ETS1) on DLK1 gene expression. The electrophoretic mobility shift assay and chromatin immunoprecipitation assay, both
and
, were used to verify the binding of the ETS1 transcription factor to the DLK1 promoter as well as the influence of methylation status of DLK1 promoter on this binding affinity. The expression of DLK1 in the right ventricular outflow tract was significantly lower in patients with Tetralogy of Fallot (TOF) than that in controls (P<0.001). Moreover, the methylation level of CpG site 10 and CpG site 11 in the DLK1_R region was significantly decreased in TOF cases compared with controls (P<0.01). The integral methylation levels of DLK1_R and the methylation status of the CpG site 11 were both positively associated with DLK1 protein expression in TOF cases. ETS1 was found to inhibit DLK1 transcriptional activity by binding to the CpG site 11 and this affinity could be influenced by the methylation level of the DLK1 promoter. These findings demonstrated that the hypomethylation of the DLK1 promoter could increase the binding affinity of ETS1 transcription factor, which in turn inhibited DLK1 gene transcriptional activity and contributed to the development of TOF.</description><identifier>ISSN: 1791-2997</identifier><identifier>EISSN: 1791-3004</identifier><identifier>DOI: 10.3892/mmr.2022.12609</identifier><identifier>PMID: 35059744</identifier><language>eng</language><publisher>Greece: Spandidos Publications</publisher><subject>Affinity ; Asians - genetics ; Base Sequence ; Binding sites (Biochemistry) ; Binding Sites - genetics ; Bisulfite ; Calcium-Binding Proteins - genetics ; Calcium-Binding Proteins - metabolism ; Cardiovascular disease ; Child, Preschool ; China ; Chromatin ; Congenital diseases ; CpG islands ; CpG Islands - genetics ; Development and progression ; DNA Methylation ; Electrophoretic mobility ; Epigenetic inheritance ; Epigenetics ; ETS protein ; Ets-1 protein ; Female ; Gene expression ; Gene Expression Regulation ; Genetic aspects ; Heart ; HEK293 Cells ; Humans ; Immunohistochemistry ; Immunoprecipitation ; Infant ; Male ; Medical research ; Medicine, Experimental ; Membrane Proteins - genetics ; Membrane Proteins - metabolism ; Methylation ; Plasmids ; Preadipocyte factor 1 ; Promoter Regions, Genetic - genetics ; Protein Binding ; Proto-Oncogene Protein c-ets-1 - genetics ; Proto-Oncogene Protein c-ets-1 - metabolism ; Sequence Analysis, DNA - methods ; Signal transduction ; Tetralogy of Fallot ; Tetralogy of Fallot - ethnology ; Tetralogy of Fallot - genetics ; Tetralogy of Fallot - metabolism ; Ventricle</subject><ispartof>Molecular medicine reports, 2022-03, Vol.25 (3), Article 93</ispartof><rights>COPYRIGHT 2022 Spandidos Publications</rights><rights>Copyright Spandidos Publications UK Ltd. 2022</rights><rights>Copyright: © Tian et al. 2022</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c485t-5b03cddd9716c26ea106222128c6a161a5884cf3a474ef8b0009f7fdebe73cb93</citedby><cites>FETCH-LOGICAL-c485t-5b03cddd9716c26ea106222128c6a161a5884cf3a474ef8b0009f7fdebe73cb93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35059744$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tian, Guixiang</creatorcontrib><creatorcontrib>He, Lili</creatorcontrib><creatorcontrib>Gu, Ruoyi</creatorcontrib><creatorcontrib>Sun, Jingwei</creatorcontrib><creatorcontrib>Chen, Weicheng</creatorcontrib><creatorcontrib>Qian, Yanyan</creatorcontrib><creatorcontrib>Ma, Xiaojing</creatorcontrib><creatorcontrib>Yan, Weili</creatorcontrib><creatorcontrib>Zhao, Zhenshan</creatorcontrib><creatorcontrib>Xu, Ziqing</creatorcontrib><creatorcontrib>Suo, Meijiao</creatorcontrib><creatorcontrib>Sheng, Wei</creatorcontrib><creatorcontrib>Huang, Guoying</creatorcontrib><title>CpG site hypomethylation at ETS1‑binding region regulates DLK1 expression in Chinese patients with Tetralogy of Fallot</title><title>Molecular medicine reports</title><addtitle>Mol Med Rep</addtitle><description>Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart malformation accounting for ~10% of cases. Although the pathogenesis of TOF is complex and largely unknown, epigenetics plays a huge role, specifically DNA methylation. The protein δ like non‑canonical Notch ligand 1 (DLK1) gene encodes a non‑canonical ligand of the Notch signaling pathway, which is involved in heart development. However, the epigenetic mechanism of DLK1 in the pathogenesis of TOF is yet to be elucidated. Therefore, the present study aimed to clarify its specific mechanism. In this study, immunohistochemistry was used to detect the protein expression of DLK1 and the methylation status of the DLK1 promoter was measured via bisulfite sequencing PCR. Dual‑luciferase reporter assays were performed to examine the influence of transcription factor ETS proto‑oncogene 1 (ETS1) on DLK1 gene expression. The electrophoretic mobility shift assay and chromatin immunoprecipitation assay, both
and
, were used to verify the binding of the ETS1 transcription factor to the DLK1 promoter as well as the influence of methylation status of DLK1 promoter on this binding affinity. The expression of DLK1 in the right ventricular outflow tract was significantly lower in patients with Tetralogy of Fallot (TOF) than that in controls (P<0.001). Moreover, the methylation level of CpG site 10 and CpG site 11 in the DLK1_R region was significantly decreased in TOF cases compared with controls (P<0.01). The integral methylation levels of DLK1_R and the methylation status of the CpG site 11 were both positively associated with DLK1 protein expression in TOF cases. ETS1 was found to inhibit DLK1 transcriptional activity by binding to the CpG site 11 and this affinity could be influenced by the methylation level of the DLK1 promoter. These findings demonstrated that the hypomethylation of the DLK1 promoter could increase the binding affinity of ETS1 transcription factor, which in turn inhibited DLK1 gene transcriptional activity and contributed to the development of TOF.</description><subject>Affinity</subject><subject>Asians - genetics</subject><subject>Base Sequence</subject><subject>Binding sites (Biochemistry)</subject><subject>Binding Sites - genetics</subject><subject>Bisulfite</subject><subject>Calcium-Binding Proteins - genetics</subject><subject>Calcium-Binding Proteins - metabolism</subject><subject>Cardiovascular disease</subject><subject>Child, Preschool</subject><subject>China</subject><subject>Chromatin</subject><subject>Congenital diseases</subject><subject>CpG islands</subject><subject>CpG Islands - genetics</subject><subject>Development and progression</subject><subject>DNA Methylation</subject><subject>Electrophoretic mobility</subject><subject>Epigenetic inheritance</subject><subject>Epigenetics</subject><subject>ETS protein</subject><subject>Ets-1 protein</subject><subject>Female</subject><subject>Gene expression</subject><subject>Gene Expression Regulation</subject><subject>Genetic aspects</subject><subject>Heart</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>Immunoprecipitation</subject><subject>Infant</subject><subject>Male</subject><subject>Medical research</subject><subject>Medicine, Experimental</subject><subject>Membrane Proteins - genetics</subject><subject>Membrane Proteins - metabolism</subject><subject>Methylation</subject><subject>Plasmids</subject><subject>Preadipocyte factor 1</subject><subject>Promoter Regions, Genetic - genetics</subject><subject>Protein Binding</subject><subject>Proto-Oncogene Protein c-ets-1 - genetics</subject><subject>Proto-Oncogene Protein c-ets-1 - metabolism</subject><subject>Sequence Analysis, DNA - methods</subject><subject>Signal transduction</subject><subject>Tetralogy of Fallot</subject><subject>Tetralogy of Fallot - ethnology</subject><subject>Tetralogy of Fallot - genetics</subject><subject>Tetralogy of Fallot - metabolism</subject><subject>Ventricle</subject><issn>1791-2997</issn><issn>1791-3004</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNptkk1v1DAQhiMEoqVw5YgsceGyi7_i2BekamkLYiUOLGfLcSZZV4kdYge6N_4Cf5FfggNL-VDlw1ieZ96Zsd6ieErwmklFXw7DtKaY0jWhAqt7xSmpFFkxjPn9450qVZ0Uj2K8xliUtFQPixNW4lJVnJ8WN5vxCkWXAO0PYxgg7Q-9SS54ZBK62H0g379-q51vnO_QBN2SyGHODET0evuOILgZJ4hxyTiPNnvnIQIaswj4FNEXl_ZoB2kyfegOKLTo0vR9SI-LB63pIzw5xrPi4-XFbvNmtX1_9XZzvl1ZLsu0KmvMbNM0qiLCUgGGYEEpJVRaYYggppSS25YZXnFoZY0xVm3VNlBDxWyt2Fnx6pfuONcDNDYPlUfR4-QGMx10ME7_m_Fur7vwWUuJFeaLwIujwBQ-zRCTHly00PfGQ5ijpss8kpGSZfT5f-h1mCef11uoivFSSPGH6kwP2vk25L52EdXnQpVMcMyrTK3voPJpYHA2eGhdfr-rwE4hxgna2x0J1otXdPaKXryif3olFzz7-2du8d_mYD8Aq4W7lQ</recordid><startdate>20220301</startdate><enddate>20220301</enddate><creator>Tian, Guixiang</creator><creator>He, Lili</creator><creator>Gu, Ruoyi</creator><creator>Sun, Jingwei</creator><creator>Chen, Weicheng</creator><creator>Qian, Yanyan</creator><creator>Ma, Xiaojing</creator><creator>Yan, Weili</creator><creator>Zhao, Zhenshan</creator><creator>Xu, Ziqing</creator><creator>Suo, Meijiao</creator><creator>Sheng, Wei</creator><creator>Huang, Guoying</creator><general>Spandidos Publications</general><general>Spandidos Publications UK Ltd</general><general>D.A. Spandidos</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AN0</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20220301</creationdate><title>CpG site hypomethylation at ETS1‑binding region regulates DLK1 expression in Chinese patients with Tetralogy of Fallot</title><author>Tian, Guixiang ; He, Lili ; Gu, Ruoyi ; Sun, Jingwei ; Chen, Weicheng ; Qian, Yanyan ; Ma, Xiaojing ; Yan, Weili ; Zhao, Zhenshan ; Xu, Ziqing ; Suo, Meijiao ; Sheng, Wei ; Huang, Guoying</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c485t-5b03cddd9716c26ea106222128c6a161a5884cf3a474ef8b0009f7fdebe73cb93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Affinity</topic><topic>Asians - genetics</topic><topic>Base Sequence</topic><topic>Binding sites (Biochemistry)</topic><topic>Binding Sites - genetics</topic><topic>Bisulfite</topic><topic>Calcium-Binding Proteins - genetics</topic><topic>Calcium-Binding Proteins - metabolism</topic><topic>Cardiovascular disease</topic><topic>Child, Preschool</topic><topic>China</topic><topic>Chromatin</topic><topic>Congenital diseases</topic><topic>CpG islands</topic><topic>CpG Islands - genetics</topic><topic>Development and progression</topic><topic>DNA Methylation</topic><topic>Electrophoretic mobility</topic><topic>Epigenetic inheritance</topic><topic>Epigenetics</topic><topic>ETS protein</topic><topic>Ets-1 protein</topic><topic>Female</topic><topic>Gene expression</topic><topic>Gene Expression Regulation</topic><topic>Genetic aspects</topic><topic>Heart</topic><topic>HEK293 Cells</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>Immunoprecipitation</topic><topic>Infant</topic><topic>Male</topic><topic>Medical research</topic><topic>Medicine, Experimental</topic><topic>Membrane Proteins - genetics</topic><topic>Membrane Proteins - metabolism</topic><topic>Methylation</topic><topic>Plasmids</topic><topic>Preadipocyte factor 1</topic><topic>Promoter Regions, Genetic - genetics</topic><topic>Protein Binding</topic><topic>Proto-Oncogene Protein c-ets-1 - genetics</topic><topic>Proto-Oncogene Protein c-ets-1 - metabolism</topic><topic>Sequence Analysis, DNA - methods</topic><topic>Signal transduction</topic><topic>Tetralogy of Fallot</topic><topic>Tetralogy of Fallot - ethnology</topic><topic>Tetralogy of Fallot - genetics</topic><topic>Tetralogy of Fallot - metabolism</topic><topic>Ventricle</topic><toplevel>online_resources</toplevel><creatorcontrib>Tian, Guixiang</creatorcontrib><creatorcontrib>He, Lili</creatorcontrib><creatorcontrib>Gu, Ruoyi</creatorcontrib><creatorcontrib>Sun, Jingwei</creatorcontrib><creatorcontrib>Chen, Weicheng</creatorcontrib><creatorcontrib>Qian, Yanyan</creatorcontrib><creatorcontrib>Ma, Xiaojing</creatorcontrib><creatorcontrib>Yan, Weili</creatorcontrib><creatorcontrib>Zhao, Zhenshan</creatorcontrib><creatorcontrib>Xu, Ziqing</creatorcontrib><creatorcontrib>Suo, Meijiao</creatorcontrib><creatorcontrib>Sheng, Wei</creatorcontrib><creatorcontrib>Huang, Guoying</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central UK/Ireland</collection><collection>British Nursing Database</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biological Sciences</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Molecular medicine reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tian, Guixiang</au><au>He, Lili</au><au>Gu, Ruoyi</au><au>Sun, Jingwei</au><au>Chen, Weicheng</au><au>Qian, Yanyan</au><au>Ma, Xiaojing</au><au>Yan, Weili</au><au>Zhao, Zhenshan</au><au>Xu, Ziqing</au><au>Suo, Meijiao</au><au>Sheng, Wei</au><au>Huang, Guoying</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>CpG site hypomethylation at ETS1‑binding region regulates DLK1 expression in Chinese patients with Tetralogy of Fallot</atitle><jtitle>Molecular medicine reports</jtitle><addtitle>Mol Med Rep</addtitle><date>2022-03-01</date><risdate>2022</risdate><volume>25</volume><issue>3</issue><artnum>93</artnum><issn>1791-2997</issn><eissn>1791-3004</eissn><abstract>Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart malformation accounting for ~10% of cases. Although the pathogenesis of TOF is complex and largely unknown, epigenetics plays a huge role, specifically DNA methylation. The protein δ like non‑canonical Notch ligand 1 (DLK1) gene encodes a non‑canonical ligand of the Notch signaling pathway, which is involved in heart development. However, the epigenetic mechanism of DLK1 in the pathogenesis of TOF is yet to be elucidated. Therefore, the present study aimed to clarify its specific mechanism. In this study, immunohistochemistry was used to detect the protein expression of DLK1 and the methylation status of the DLK1 promoter was measured via bisulfite sequencing PCR. Dual‑luciferase reporter assays were performed to examine the influence of transcription factor ETS proto‑oncogene 1 (ETS1) on DLK1 gene expression. The electrophoretic mobility shift assay and chromatin immunoprecipitation assay, both
and
, were used to verify the binding of the ETS1 transcription factor to the DLK1 promoter as well as the influence of methylation status of DLK1 promoter on this binding affinity. The expression of DLK1 in the right ventricular outflow tract was significantly lower in patients with Tetralogy of Fallot (TOF) than that in controls (P<0.001). Moreover, the methylation level of CpG site 10 and CpG site 11 in the DLK1_R region was significantly decreased in TOF cases compared with controls (P<0.01). The integral methylation levels of DLK1_R and the methylation status of the CpG site 11 were both positively associated with DLK1 protein expression in TOF cases. ETS1 was found to inhibit DLK1 transcriptional activity by binding to the CpG site 11 and this affinity could be influenced by the methylation level of the DLK1 promoter. These findings demonstrated that the hypomethylation of the DLK1 promoter could increase the binding affinity of ETS1 transcription factor, which in turn inhibited DLK1 gene transcriptional activity and contributed to the development of TOF.</abstract><cop>Greece</cop><pub>Spandidos Publications</pub><pmid>35059744</pmid><doi>10.3892/mmr.2022.12609</doi><oa>free_for_read</oa></addata></record> |
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| ispartof | Molecular medicine reports, 2022-03, Vol.25 (3), Article 93 |
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| subjects | Affinity Asians - genetics Base Sequence Binding sites (Biochemistry) Binding Sites - genetics Bisulfite Calcium-Binding Proteins - genetics Calcium-Binding Proteins - metabolism Cardiovascular disease Child, Preschool China Chromatin Congenital diseases CpG islands CpG Islands - genetics Development and progression DNA Methylation Electrophoretic mobility Epigenetic inheritance Epigenetics ETS protein Ets-1 protein Female Gene expression Gene Expression Regulation Genetic aspects Heart HEK293 Cells Humans Immunohistochemistry Immunoprecipitation Infant Male Medical research Medicine, Experimental Membrane Proteins - genetics Membrane Proteins - metabolism Methylation Plasmids Preadipocyte factor 1 Promoter Regions, Genetic - genetics Protein Binding Proto-Oncogene Protein c-ets-1 - genetics Proto-Oncogene Protein c-ets-1 - metabolism Sequence Analysis, DNA - methods Signal transduction Tetralogy of Fallot Tetralogy of Fallot - ethnology Tetralogy of Fallot - genetics Tetralogy of Fallot - metabolism Ventricle |
| title | CpG site hypomethylation at ETS1‑binding region regulates DLK1 expression in Chinese patients with Tetralogy of Fallot |
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