title: Hedgehog signaling through GLI1 and GLI2 is required for epithelial-mesenchymal transition in human trophoblasts.
abstract: BACKGROUND
  Epithelial to mesenchymal transition (EMT) is critical for human placental development, trophoblastic differentiation, and pregnancy-associated diseases. Here, we investigated the effects of hedgehog (HH) signaling on EMT in human trophoblasts, and further explored the underlying mechanism.
  
  
  METHODS
  Human primary cytotrophoblasts and trophoblast-like JEG-3 cells were used as in vitro models. Quantitative real-time RT-PCR and Western blot analysis were performed to examine mRNA and protein levels, respectively. Lentiviruses expressing short hairpin RNA were used to knock down the target genes. Reporter assays and chromatin immunoprecipitation were performed to determine the transactivity. Cell migration, invasion and colony formation were accessed by wound healing, Matrigel-coated transwell, and colony formation assays, respectively.
  
  
  RESULTS
  Activation of HH signaling induced the transdifferentiation of cytotrophoblasts and trophoblast-like JEG-3 cells from epithelial to mesenchymal phenotypes, exhibiting the decreases in E-Cadherin expression as well as the increases in vimentin expression, invasion, migration and colony formation. Knockdown of GLI1 and GLI2 but not GLI3 attenuated HH-induced transdifferentiation, whereas GLI1 was responsible for the expression of HH-induced key EMT regulators including Snail1, Slug, and Twist, and both GLI1 and GLI2 acted directly as transcriptional repressor of CDH1 gene encoding E-Cadherin.
  
  
  CONCLUSIONS
  HH through GLI1 and GLI2 acts as critical signals in supporting the physiological function of mature placenta.
  
  
  CONCLUSIONS
  HH signaling through GLI1 and GLI2 could be required for the maintenance of human pregnancy.
authors.affiliation: Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Tang
authors.affiliation: Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Mei
authors.affiliation: Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Liyu
authors.initials: L
authors.lastname: Pan
authors.affiliation: The Affiliated Children Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Wenyi
authors.initials: W
authors.lastname: Xiong
authors.affiliation: The Affiliated Children Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Haibin
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Hongfeng
authors.initials: H
authors.lastname: Ruan
authors.affiliation: The Affiliated Children Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Chaochun
authors.initials: C
authors.lastname: Zou
authors.affiliation: The Affiliated Children Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Lanfang
authors.initials: L
authors.lastname: Tang
authors.affiliation: Department of Toxicology, Osaka University, Suita, Osaka, Japan.
authors.forename: Takuma
authors.initials: T
authors.lastname: Iguchi
authors.affiliation: Department of Pharmacology, School of Medicine, Zhejiang University, Hangzhou, China. Electronic address: xiwu@zju.edu.cn.
authors.forename: Ximei
authors.initials: X
authors.lastname: Wu
mesh_terms: D015153:Blotting, Western; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D058750:Epithelial-Mesenchymal Transition; D005260:Female; D015870:Gene Expression; D057809:HEK293 Cells; D053823:Hedgehog Proteins / Q000378:metabolism*; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D008954:Models, Biological; D010920:Placenta / Q000166:cytology / Q000378:metabolism; D011247:Pregnancy; D011485:Protein Binding; D034622:RNA Interference; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D000072086:Smoothened Receptor; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism*; D000071676:Zinc Finger Protein GLI1; D000075746:Zinc Finger Protein Gli2
chemical_list: D015820:Cadherins; C400551:GLI1 protein, human; D053823:Hedgehog Proteins; D051741:Kruppel-Like Transcription Factors; D043562:Receptors, G-Protein-Coupled; C491023:SMO protein, human; D000072086:Smoothened Receptor; D014157:Transcription Factors; D000071676:Zinc Finger Protein GLI1; D000075746:Zinc Finger Protein Gli2
keywords: Epithelial–mesenchymal transition; Hedgehog signaling; Trophoblast