Hedgehog signaling through GLI1 and GLI2 is required for epithelial–mesenchymal transition in human trophoblasts

• Published in: Biochimica et Biophysica Acta (BBA) - General Subjects Vol. 1850; no. 7; pp. 1438 - 1448
• Main Authors: Tang, Chao, Mei, Liu, Pan, Liyu, Xiong, Wenyi, Zhu, Haibin, Ruan, Hongfeng, Zou, Chaochun, Tang, Lanfang, Iguchi, Takuma, Wu, Ximei
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2015; Elsevier BV
• Subjects: Blotting, Western; Cadherins; Cadherins - genetics; Cadherins - metabolism; Cell Line; cell movement; Cells, Cultured; chromatin; Epithelial-Mesenchymal Transition; epithelium; Female; Gene Expression; genes; Hedgehog Proteins; Hedgehog Proteins - metabolism; Hedgehog signaling; HEK293 Cells; Humans; Kruppel-Like Transcription Factors; Kruppel-Like Transcription Factors - genetics; Kruppel-Like Transcription Factors - metabolism; Lentivirus; messenger RNA; Microscopy, Confocal; Models, Biological; phenotype; Placenta; Placenta - cytology; Placenta - metabolism; precipitin tests; Pregnancy; Protein Binding; quantitative polymerase chain reaction; Receptors, G-Protein-Coupled; Receptors, G-Protein-Coupled - genetics; Receptors, G-Protein-Coupled - metabolism; repressor proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Signal Transduction; small interfering RNA; Smoothened Receptor; tissue repair; Transcription Factors; Transcription Factors - genetics; Transcription Factors - metabolism; Trophoblast; Trophoblasts; Trophoblasts - cytology; Trophoblasts - metabolism; vimentin; Western blotting; Zinc Finger Protein GLI1; Zinc Finger Protein Gli2; Hedgehog signaling; Epithelial–mesenchymal transition; Trophoblast
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2015.04.005

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. 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. 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. HH through GLI1 and GLI2 acts as critical signals in supporting the physiological function of mature placenta. HH signaling through GLI1 and GLI2 could be required for the maintenance of human pregnancy. •Activation of hedgehog signaling induces EMT in human trophoblasts.•GLI1 and GLI2 mediate hedgehog-induced transcriptional suppression of CDH1 gene.•GLI1 alone mediates hedgehog-induced transcription of key EMT regulators.•Hedgehog signaling is possibly critical for pregnancy-associated diseases.