The YY1/MMP2 axis promotes trophoblast invasion at the maternal-fetal interface
YY1 is a sequence‐specific DNA‐binding transcription factor that has many important biological roles. However, its function in trophoblasts at the maternal–fetal interface remains to be elucidated. In this study, we used an mRNA microarray and reverse transcription qPCR and compared the YY1 mRNA exp...
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| Published in | The Journal of pathology Vol. 239; no. 1; pp. 36 - 47 |
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| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Chichester, UK
John Wiley & Sons, Ltd
01.05.2016
Wiley Subscription Services, Inc |
| Subjects | |
| Online Access | Get full text |
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| Summary: | YY1 is a sequence‐specific DNA‐binding transcription factor that has many important biological roles. However, its function in trophoblasts at the maternal–fetal interface remains to be elucidated. In this study, we used an mRNA microarray and reverse transcription qPCR and compared the YY1 mRNA expression level in trophoblasts between patients with recurrent miscarriage (RM) and healthy control subjects. Our results revealed that YY1 mRNA expression was significantly lower in the trophoblasts of the RM group compared with the healthy control group. Furthermore, immunofluorescence and immunohistochemical data showed that YY1 was highly expressed in human placental villi during early pregnancy, especially in cytotrophoblast cells and invasive extravillous trophoblasts, and it was expressed at a much lower level in the placental villi of term pregnancy. YY1 overexpression enhanced, and knockdown repressed, the invasion and proliferation of trophoblasts. Antibody array screening revealed that YY1 significantly promoted MMP2 expression in trophoblasts. Bioinformatics analysis identified three YY1‐binding sites in the MMP2 promoter region, and chromatin immunoprecipitation analysis verified that YY1 binds directly to its promoter region. Importantly, inhibition of YY1 by siRNA clearly decreased trophoblast invasion in an ex vivo explant culture model. Overall, our findings revealed a new regulatory pathway of YY1/MMP2 in trophoblast cell invasion during early pregnancy and indicated that YY1 may be involved in the pathogenesis of RM. © 2016 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland. |
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| Bibliography: | Shanghai Jiao Tong University Medicine-Engineering Fund - No. YG2013ZD04 ArticleID:PATH4694 AppendixS1. SUPPORTING INFORMATIONNetwork of YY1 interacting proteins, as determined by the STRING (Search Tool for the Retrieval of Interacting Genes/Proteins) database.YY1 promoted trophoblast migration. (A and B) Overexpression of YY1 in HTR-8 cells resulted in increased wound closure ability compared with the vector control cells. YY1 knockdown drastically reduced the rate of wound closure in comparison to the scrambled control cell line (magnification × 100). The migration distance of trophoblast was quantified using Imagepro-plus 6.0 software (p < 0.05 compared with siCtrl or vector).YY1 promoted trophoblast migration from extravillous explants. (A) Extravillous explants were maintained in culture on Matrigel. Serial pictures of the explants incubated with siYY1 or siCtrl were taken under a light microscope after 24 h and 72 h of culture in vitro. A statistical analysis of the migration distance of villous tips (%) is shown. (B) Extravillous explants from RM patients were maintained in culture on Matrigel. Serial pictures of the explants incubated with lenti-ctrl or lenti-YY1 lentivirus were taken under a light microscope after 24 h and 72 h of culture in vitro. A statistical analysis of the migration distance of villous tips (%) is shown (p < 0.05 compared with siCtrl or lenti-ctrl).YY1 promoted MMP2 expression in trophoblasts. (A-D) HTR-8 cells were transfected with siCtrl, siYY1, and vector or YY1 overexpression vector for 48 h. The MMP2 expression level was quantified using mean fluorescent intensities. Fluorescence intensity of the signals was assessed using Leica confocal SP 8 software (p < 0.05 compared with siCtrl or vector).Level of MMP2 in villous sections from RM patients was significantly decreased. (A and B): Representative immunofluorescence of MMP2 in frozen first-trimester villous sections obtained at 6-8 weeks of gestation. Fluorescence signals specific to anti-YY1 antibodies are green and the DAPI-stained nuclei are blue. Fluorescence intensity of the signals was assessed using Leica confocal SP 8 software (p < 0.05 compared with RM.YY1 siRNA inhibits EVT migration by down-regulating MMP2. (A and B) Extravillous explants (6-8 weeks of gestation) were maintained in culture on Matrigel for 24 h, and transfected with siYY1 and siCtrl for 48 h. Fluorescence signals specific to anti-YY1 antibodies appear green; CK7, red; and DAPI-stained nuclei, blue. (C and D) Fluorescence intensity of the signals was assessed using Leica confocal SP 8 software (p < 0.05 compared with siCtrl). Shanghai Natural Science Fund Project - No. 14ZR1443800 National Basic Research Program of China - No. 2013CB967404 National Natural Science Foundation of China - No. 81401218; No. 81370767; No. 81501250; No. 81125004; No. 31171439 Joint Key Project of New Frontier Technology in Shanghai Municipal Hospitals, Shanghai Municipal - No. SHDC12014129 istex:9425D3DAAE209000F208AB4C1FCBA87E2948AD35 Fund for Outstanding Academic Leaders in Shanghai, China - No. 12XD1406600 ark:/67375/WNG-DR6582PC-R ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 These authors contributed equally to this work. No conflicts of interest were declared |
| ISSN: | 0022-3417 1096-9896 |
| DOI: | 10.1002/path.4694 |