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LncRNA NEAT1 Promotes TLR4 Expression to Regulate Lipopolysaccharide-Induced Trophoblastic Cell Pyroptosis as a Molecular Sponge of miR-302b-3p

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Abstract

Pyroptosis is an inflammation-triggered cell death caused by certain inflammasomes, and long non-coding RNAs (lncRNAs) are related to cell pyroptosis. This study evaluated the mechanism of lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) on lipopolysaccharide (LPS)-induced trophoblastic cells pyroptosis. HTR-8/Svneo trophoblastic cells were treated with LPS. The expression of lncRNA NEAT1 was decreased using siRNAs, followed by the evaluation of cell proliferation, Caspase-1 activity, levels of Cleaved Caspase-1 and gasdermin D-N, and the concentrations of Interleukin (IL)-1β and IL-18. We found that LPS promoted the pyroptosis of HTR-8/Svneo cells, and lncRNA NEAT1 was highly expressed in LPS-treated HTR-8/Svneo cells while silencing lncRNA NEAT1 inhibited LPS-induced trophoblastic cells pyroptosis. The subcellular localization of lncRNA NEAT1 was detected. Dual-luciferase gene experiment and RNA pull-down assay detected that lncRNA NEAT1 bound to miR-302b-3p and could inhibit miR-302b-3p, and toll-like receptor 4 (TLR4) was the target gene of miR-302b-3p. Then, a joint experiment was designed for detection, which found that miR-302b-3p downregulation partially reversed the inhibition of silencing lncRNA NEAT1 on LPS-induced trophoblastic cells pyroptosis and overexpression of TLR4 annulled the inhibition of silencing lncRNA NEAT1 on LPS-induced trophoblastic cells pyroptosis. Therefore, lncRNA NEAT1 promoted the transcription of TLR4 by competitively binding to miR-302b-3p, thus promoting LPS-induced trophoblastic cells pyroptosis.

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Data Availability

The data that support this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank all of members in our team for the excellent work.

Funding

This work was supported by Research Project of Maternal and Child Health in Jiangsu Province under Grant Number F201944.

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Authors and Affiliations

  1. Department of Prenatal Diagnosis, Clinical Medical College, Northern Jiangsu People’s Hospital, Yangzhou University, No. 98 Nantong West Road, Guangling District, Yangzhou, Jiangsu Province, 225001, China

    Dan Fu, Yun Ju & Suhua Zhang

  2. Reproductive Medicine Center, Clinical Medical College, Northern Jiangsu People’s Hospital, Yangzhou University, No. 98 Nantong West Road, Guangling District, Yangzhou, Jiangsu Province, 225001, China

    Chunhui Zhu & Yu Pan

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  1. Dan Fu
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  2. Yun Ju
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  5. Suhua Zhang
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Contributions

Conceptualization: DF, YJ, and CZ; data curation: DF and Yu Pan; methodology: DF, YJ, and SZ; validation: CZ and Yu Pan; writing—original draft: DF and YJ; writing—review and editing: CZ and SZ.

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Correspondence to Suhua Zhang.

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Fu, D., Ju, Y., Zhu, C. et al. LncRNA NEAT1 Promotes TLR4 Expression to Regulate Lipopolysaccharide-Induced Trophoblastic Cell Pyroptosis as a Molecular Sponge of miR-302b-3p. Mol Biotechnol 64, 670–680 (2022). https://doi.org/10.1007/s12033-021-00436-2

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