title: The effect of cigarette smoke extract on trophoblast cell viability and migration: the role of adrenomedullin.
abstract: We tested the hypothesis that cigarette smoke extract (CSE) leads to differences in expression of genes involved in angiogenesis and affects cell viability and migration in a first-trimester cytotrophoblast cell line (HTR-8/SVneo). HTR-8/SVneo cells were treated with 1% CSE, and gene expression for adrenomedullin (ADM), placental growth factor (PlGF), soluble fms-like tyrosine kinase 1 (sFLT-1), and vascular endothelia growth factor (VEGF) and protein content for ADM, PlGF, and sFlt-1 determined. A cell viability assay and a cell migration scratch assay were utilized following treatment with CSE with and without ADM inhibitor. Adrenomedullin, PlGF, and VEGF gene transcripts were significantly upregulated by 1% CSE treatment compared with unstimulated cells or cells treated with nicotine alone. Neither 1% CSE nor nicotine treatment alone affected sFlt-1 gene expression. There was a significant increase in secreted ADM protein from cells treated with 1% CSE detected by enzyme-linked immunosorbant assay, though no differences in PlGF or sFlt-1 production were seen. Treatment with 1% CSE increased cell viability and cell migration compared with unstimulated cells and was inhibited by co-treatment with ADM inhibitor. Treatment of a first-trimester trophoblast cell line with CSE increases cell viability and cell migration that are reversed by co-treatment with ADM inhibitor, suggesting that ADM at least partially mediates cell growth and viability following CSE treatment.
authors.affiliation: Division of Maternal-Fetal Medicine, Department of Obstetrics and Gynecology, Duke University School of Medicine, Durham, NC 27710, USA.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Beiswenger
authors.forename: Liping
authors.initials: L
authors.lastname: Feng
authors.forename: Haywood L
authors.initials: HL
authors.lastname: Brown
authors.forename: R Phillips
authors.initials: RP
authors.lastname: Heine
authors.forename: Amy P
authors.initials: AP
authors.lastname: Murtha
authors.forename: Chad A
authors.initials: CA
authors.lastname: Grotegut
mesh_terms: D053607:Adrenomedullin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D009538:Nicotine / Q000494:pharmacology; D000072483:Placenta Growth Factor; D011225:Pre-Eclampsia; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000235:genetics; D011261:Pregnancy Trimester, First; D012906:Smoke / Q000009:adverse effects / Q000032:analysis; D014026:Nicotiana; D014327:Trophoblasts / Q000166:cytology* / Q000187:drug effects; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics
chemical_list: C000608036:PGF protein, human; D011257:Pregnancy Proteins; D012906:Smoke; D042461:Vascular Endothelial Growth Factor A; D000072483:Placenta Growth Factor; D053607:Adrenomedullin; D009538:Nicotine; D040281:Vascular Endothelial Growth Factor Receptor-1