Journal of Shanghai Jiao Tong University (Medical Science) >

2022 , Vol. 42 >Issue 9: 1247 - 1257

DOI: https://doi.org/10.3969/j.issn.1674-8115.2022.09.011

Basic research

TCF3 knockdown inhibits the decidualization of human endometrial stromal cells

  • Xiaowei WEI ,
  • Fuju TIAN ,
  • Xiaorui LIU ,
  • Weihong ZENG ,
  • Cailian CHEN ,
  • Yi LIN
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  • 1.Central Laboratory of The International Peace Maternity & Child Health Hospital, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Embryo Original Diseases; Institute of Birth Defects and Rare Diseases, Shanghai Jiao Tong University School of Medicine, Shanghai 200030, China
    2.Department of Automation, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University; Key Laboratory of System Control and Information Processing, Ministry of Education, Shanghai 200240, China
LIN Yi, E-mail: yilinonline@126.com.

Received date: 2022-05-19

  Accepted date: 2022-08-29

  Online published: 2022-09-28

Supported by

National Key Research and Development Program of China(2018YFC1002800);National Natural Science Foundation of China(82171669);Shanghai Jiao Tong University Trans-Med Awards Research (Major Project)(20210201)

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Abstract

Objective ·To investigate the effect of transcription factor 3 (TCF3) on decidualization of human endometrial stromal cells (HESCs). Methods ·HESC decidualization in vitro was induced by 8-bromoadenosine 3',5'-cyclic adenosine monophosphate (8-Br-cAMP) and medroxy progesterone acetate (MPA). Small interfering RNA (siRNA) against TCF3 was used to construct TCF3-knockdown cell model. Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting were used to confirm the knockdown efficiency of TCF3. RT-qPCR and enzyme-linked immunosorbent assay (ELISA) were used to detect the expression of decidual markers, including insulin like growth factor binding protein 1 (IGFBP1) and prolactin (PRL). The morphological changes of HESCs after decidualization and after TCF3 knockdown were detected by Alexa Fluor-labeled phalloidin staining. The HESCs were divided into three groups: non-decidualized+NC (transfected with control siRNA) group, decidualized 4 d+NC group, and decidualized 4 d+siTCF3 (transfected with TCF3 siRNA) group, and RNA sequencing was performed. In order to explore the mechanism of TCF3 regulating the decidualization of HESCs, cluster analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, and gene set enrichment analysis (GSEA) were conducted in the three groups of cells. qPCR was used to confirm the results of RNA sequencing analysis. Results ·The expression level of TCF3 increased gradually after 2 d and 4 d of decidualization of HESCs. Both ELISA and RT-qPCR results showed that after decidualization for 2 d or 4 d, the expression of decidual markers IGFBP1 and PRL decreased after TCF3 knockdown. Phalloidin staining results showed that after decidualization, the cell morphology changed from a long and slender fusiform structure to a large and round cell shape, while after TCF3 was knockdown, the cell morphology returned to the long fusiform structure. RNA sequencing analysis showed that the differential genes were mainly enriched in the cytokine-cytokine receptor pathway, which was further verified by GSEA analysis. RT-qPCR results confirmed that knockdown of TCF3 down-regulated the expression of several cytokines, such as leukemia inhibitory factor (LIF), interleukin 6 (IL-6), interleukin 1 β (IL1B), interleukin 1 receptor type 1 (IL1R1), and interleukin 1 receptor type 1 (CSF1). Conclusion ·Down-regulation of TCF3 in HESCs inhibits decidualization maybe through regulating the expression of cytokines IL-6, IL1B, IL1R1 and CSF1.

Key words： human endometrial stromal cell (HESC); transcription factor 3 (TCF3); decidualization; cytokine

Cite this article

Xiaowei WEI , Fuju TIAN , Xiaorui LIU , Weihong ZENG , Cailian CHEN , Yi LIN . TCF3 knockdown inhibits the decidualization of human endometrial stromal cells[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022 , 42(9) : 1247 -1257 . DOI: 10.3969/j.issn.1674-8115.2022.09.011

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