title: Transcriptome analysis of PPARγ target genes reveals the involvement of lysyl oxidase in human placental cytotrophoblast invasion.
abstract: Human placental development is characterized by invasion of extravillous cytotrophoblasts (EVCTs) into the uterine wall during the first trimester of pregnancy. Peroxisome proliferator-activated receptor γ (PPARγ) plays a major role in placental development, and activation of PPARγ by its agonists results in inhibition of EVCT invasion in vitro. To identify PPARγ target genes, microarray analysis was performed using GeneChip technology on EVCT primary cultures obtained from first-trimester human placentas. Gene expression was compared in EVCTs treated with the PPARγ agonist rosiglitazone versus control. A total of 139 differentially regulated genes were identified, and changes in the expression of the following 8 genes were confirmed by reverse transcription-quantitative polymerase chain reaction: a disintegrin and metalloproteinase domain12 (ADAM12), connexin 43 (CX43), deleted in liver cancer 1 (DLC1), dipeptidyl peptidase 4 (DPP4), heme oxygenase 1 (HMOX-1), lysyl oxidase (LOX), plasminogen activator inhibitor 1 (PAI-1) and PPARγ. Among the upregulated genes, lysyl oxidase (LOX) was further analyzed. In the LOX family, only LOX, LOXL1 and LOXL2 mRNA expression was significantly upregulated in rosiglitazone-treated EVCTs. RNA and protein expression of the subfamily members LOX, LOXL1 and LOXL2 were analyzed by absolute RT-qPCR and western blotting, and localized by immunohistochemistry and immunofluorescence-confocal microscopy. LOX protein was immunodetected in the EVCT cytoplasm, while LOXL1 was found in the nucleus and nucleolus. No signal was detected for LOXL2 protein. Specific inhibition of LOX activity by β-aminopropionitrile in cell invasion assays led to an increase in EVCT invasiveness. These results suggest that LOX, LOXL1 and LOXL2 are downstream PPARγ targets and that LOX activity is a negative regulator of trophoblastic cell invasion.
authors.affiliation: INSERM, UMR-S767, Paris, France ; Université Paris Descartes, Sorbonne Paris Cité, Paris, France ; PremUP Foundation, Paris, France.
authors.forename: Nadine
authors.initials: N
authors.lastname: Segond
authors.forename: Séverine A
authors.initials: SA
authors.lastname: Degrelle
authors.forename: Sarah
authors.initials: S
authors.lastname: Berndt
authors.forename: Elodie
authors.initials: E
authors.lastname: Clouqueur
authors.forename: Christine
authors.initials: C
authors.lastname: Rouault
authors.forename: Bruno
authors.initials: B
authors.lastname: Saubamea
authors.forename: Philippe
authors.initials: P
authors.lastname: Dessen
authors.forename: Keith S K
authors.initials: KS
authors.lastname: Fong
authors.forename: Katalin
authors.initials: K
authors.lastname: Csiszar
authors.forename: Josette
authors.initials: J
authors.lastname: Badet
authors.forename: Danièle
authors.initials: D
authors.lastname: Evain-Brion
authors.forename: Thierry
authors.initials: T
authors.lastname: Fournier
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000235:genetics / Q000378:metabolism; D000629:Aminopropionitrile / Q000494:pharmacology; D005260:Female; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006801:Humans; D047495:PPAR gamma / Q000378:metabolism*; D010920:Placenta / Q000187:drug effects / Q000201:enzymology; D010929:Placentation; D011247:Pregnancy; D011261:Pregnancy Trimester, First / Q000235:genetics / Q000502:physiology; D021381:Protein Transport / Q000187:drug effects; D008249:Protein-Lysine 6-Oxidase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D000077154:Rosiglitazone; D045162:Thiazolidinediones / Q000494:pharmacology; D014327:Trophoblasts / Q000166:cytology* / Q000187:drug effects
chemical_list: D047495:PPAR gamma; D045162:Thiazolidinediones; D000077154:Rosiglitazone; D000629:Aminopropionitrile; D000594:Amino Acid Oxidoreductases; C445313:LOXL1 protein, human; C119091:LOXL2 protein, human; D008249:Protein-Lysine 6-Oxidase