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During placentation, epithelial cytotrophoblast cells (CTBs) transdifferentiate into migratory extravillous trophoblasts (EVTs) that invade the maternal endometrium. Improper differentiation can result in inadequate EVT invasion leading to placental malformation-related pathologies like preeclampsia. Acquisition of invasive phenotype by EVTs indicates the involvement of epithelial to mesenchymal transition (EMT). In the early-stage placenta, the trophoblast cells are exposed to a hypoxic environment. However, the regulation of EMT in trophoblasts by this hypoxic condition is unclear. Therefore, we analyzed the expression pattern of EMT-associated genes and their DNA methylation level in two different trophoblast cell lines grown under hypoxia. Exposure to hypoxia was confirmed by a significant increase in the expression of HIF1A target gene Carbonic Anhydrase 9. Modulated expression of some EMT-associated core genes indicated the induction of EMT in trophoblasts by hypoxia. Interestingly, a significant increase in the expression of MMP2 and MMP9 was observed in HTR8\/SVneo cells but not in JEG-3 cells. In HTR8\/SVneo cells, hypoxia-induced modulation in the methylation levels of promoters for E-Cadherin and MMP9 gene correlated well with alterations in their gene expression. The expression of TET1, a DNA demethylating enzyme, also increased after hypoxia exposure. Thus, we concluded that hypoxia changes expression and promoter methylation of some EMT-associated genes in trophoblast cells.<\/jats:p>","DOI":"10.1371\/journal.pone.0325053","type":"journal-article","created":{"date-parts":[[2026,4,9]],"date-time":"2026-04-09T17:24:52Z","timestamp":1775755492000},"page":"e0325053","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Hypoxia regulates epithelial to mesenchymal transition-associated genes in human trophoblast cells by modulating DNA methylation"],"prefix":"10.1371","volume":"21","author":[{"given":"Jaganmoy","family":"Choudhury","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bodhana","family":"Dhole","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kanika","family":"Aggarwal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Palak","family":"Singh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Moses Azaraiah","family":"Jala","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deepak","family":"Pandey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pradeep 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