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Although long non\u2010coding RNAs (lncRNAs) have been implicated in heart development and several CHDs, their role in TOF is not well understood. This study aimed to investigate how dysregulated lncRNAs contribute to TOF. Using Gene Expression Omnibus data mining, bioinformatics analysis and clinical heart tissue sample detecting, we identified a novel antisense lncRNA TBX5\u2010AS1:2 with unknown function that was significantly down\u2010regulated in injured cardiac tissues from TOF patients. LncRNA TBX5\u2010AS1:2 was mainly located in the nucleus of the human embryonic kidney 293 (HEK293T) cells and formed an RNA\u2010RNA double\u2010stranded structure in the overlapping region with its sense mRNA T\u2010box transcription factor\u00a05 (<jats:italic>TBX5<\/jats:italic>), which is an important regulator in heart development. Knock\u2010down of lncRNA TBX5\u2010AS1:2 via promoter hypermethylation reduced <jats:italic>TBX5<\/jats:italic> expression at both the mRNA and protein levels by affecting its mRNA stability through RNA\u2010RNA interaction. Moreover, lncRNA TBX5\u2010AS1:2 knock\u2010down inhibited the proliferation of HEK293T cells. In conclusion, these results indicated that lncRNA TBX5\u2010AS1:2 may be involved in TOF by affecting cell proliferation by targeting <jats:italic>TBX5<\/jats:italic>.<\/jats:p>","DOI":"10.1111\/jcmm.15298","type":"journal-article","created":{"date-parts":[[2020,5,5]],"date-time":"2020-05-05T11:41:39Z","timestamp":1588678899000},"page":"6472-6484","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Hypermethylation\u2010mediated down\u2010regulation of lncRNA TBX5\u2010AS1:2 in Tetralogy of Fallot inhibits cell proliferation by reducing <i>TBX5<\/i> expression"],"prefix":"10.1111","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9074-8570","authenticated-orcid":false,"given":"Jing","family":"Ma","sequence":"first","affiliation":[{"name":"Department of Facial Plastic and Reconstructive Surgery ENT Institute Eye &amp; ENT Hospital Fudan University Shanghai China"},{"name":"Department of Biochemistry and Molecular Biology Research Center for Birth Defects Institutes of Biomedical Sciences Key Laboratory of Metabolism and Molecular Medicine Ministry of Education School of Basic Medical Sciences Fudan University Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiyu","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Research Center for Birth Defects Institutes of Biomedical Sciences Key Laboratory of Metabolism and Molecular Medicine Ministry of Education School of Basic Medical Sciences Fudan University Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7369-9276","authenticated-orcid":false,"given":"Lili","family":"Hao","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology Research Center for Birth Defects Institutes of Biomedical Sciences Key Laboratory of Metabolism and Molecular Medicine 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