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Abnormal H3K27 histone methylation of RASA1 gene leads to unexplained recurrent spontaneous abortion by regulating Ras-MAPK pathway in trophoblast cells

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Abstract

Some studies suggest that the inactivation of the Ras-MAPK pathway in trophoblast cells can lead to recurrent abortion, but the molecular mechanism underlying the inactivation of this pathway in trophoblast cells is still unclear. This study aimed to explore the relationship between the mechanism of abnormal activation of RASA1, a regulatory protein of the Ras-MAPK pathway, and unexplained recurrent spontaneous abortion. RT-qPCR was used to detect the transcription levels of RASA1 gene. Immunohistochemistry and Western blot were used to detect the expression levels of the RASA1, Raf and MEK proteins. CCK-8, TUNEL and Transwell assays were used to detect the proliferative, apoptotic, and invasive capacities of HTR-8/SVneo cells. ChIP assays were used to detect the enrichment of H3K27me3 in RASA1 gene promoter. Abortion villi experiments showed that the enrichment of H3K27me3 in the RASA1 gene promoter was reduced, and that both RASA1 gene transcription and RASA1 protein expression were increased. Cell experiments confirmed that RASA1 could decrease the phosphorylated Raf and MEK proteins, inhibit the proliferation and invasion ability, and promote the apoptosis ability of HTR-8/SVneo cells. It was also found that the proliferation and invasion ability as well as the Ras-MAPK pathway activity of HTR-8/SVneo cells were inhibited when treated with histone methyltransferase inhibitor DZNep. RASA1 gene was abnormally activated in unexplained recurrent spontaneous abortion villi due to the decreased enrichment of H3K27me3 in the gene promoter. High expression of RASA1 could inhibit the activity of the Ras-MAPK pathway, and thus inhibit the proliferation and invasion ability of trophoblast cells.

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Data Availability

The analyzed data sets generated during the present study are available from the corresponding author on reasonable request.

Abbreviations

ChIP:

Chromatin immunoprecipitation

lncRNA:

Long noncoding RNA

MEK:

Mitogen-activated protein kinase kinase

Raf:

Rapidly accelerated fibrosarcoma

RASA1:

RAS P21 Protein Activator 1

shRNA:

Short hairpin RNA

URSA:

Unexplained recurrent spontaneous abortion

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Funding

The Project Supported by The Natural Science Foundation of Guangdong Province (Grant No. 2018A0303100021), National Natural Science Foundation of China (Grant No. 81971385), National Natural Science Foundation of China (Grant No. 81902751), Natural Science Foundation of Guangdong Province (Grant No. 2019A1515010412) and Researcher Cultivation Project of Shenzhen People's Hospital (Grant No. SYKYPY201927).

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Shenzhen People’s Hospital (The Second Clinical Medical College, Jinan University), Shenzhen, 518020, People’s Republic of China

    Jun Zhang & Xinqiong Liu

  2. Department of Ophthalmology, Shenzhen People’s Hospital (The Second Clinical Medical College, Jinan University), Shenzhen, 518020, People’s Republic of China

    Yali Gao

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  1. Jun Zhang
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  2. Xinqiong Liu
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Contributions

ZJ and GYL researched conception and design. ZJ analyzed data and interpretation. GYL and LXQ analyzed statistically. ZJ and GYL drafted the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Yali Gao.

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No potential conflicts of interest were disclosed.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the ethic committee of the Shenzhen People’s Hospital.

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All patients in the present study provided written informed consent.

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All authors have read the manuscript and approved the final version.

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Zhang, J., Liu, X. & Gao, Y. Abnormal H3K27 histone methylation of RASA1 gene leads to unexplained recurrent spontaneous abortion by regulating Ras-MAPK pathway in trophoblast cells. Mol Biol Rep 48, 5109–5119 (2021). https://doi.org/10.1007/s11033-021-06507-6

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  • DOI: https://doi.org/10.1007/s11033-021-06507-6

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