Abstract
Purpose
To evaluate whether miR-148a-3p overexpression is associated with disrupted decidualization of recurrent implantation failure (RIF).
Methods
Endometrial miRNA and mRNA expression profiles during the implantation window derived from women with and without RIF were identified using microarray and RT-qPCR. Immortalized human endometrial stromal cells (HESCs) were cultured for proliferation and in vitro decidualization assays after enhancing miR-148a-3p expression or inhibiting putative target gene homeobox C8 (HOXC8) expression. RT-qPCR, western blot, and luciferase reporter assays were used to confirm the relationship between miR-148a-3p and HOXC8 gene.
Results
MiR-148a-3p was significantly upregulated in RIF endometrial tissues. Forced expression of miR-148a-3p notably attenuated HESC in vitro decidualization. Mechanistic studies revealed that miR-148a-3p directly bounds to the HOXC8 3′ untranslated region (3′UTR) and suppressed HOXC8 expressions in both mRNA and protein levels. Further investigations demonstrated that inhibition of HOXC8 in HESCs induced similar effects on decidual process as those induced by miR-148a-3p overexpression.
Conclusion
Taken together, our findings suggested that elevated miR-148a-3p might account for flawed decidualization in RIF by negatively regulating HOXC8, raising the possibility that miR-148a-3p might be a novel therapeutic target in RIF.
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Acknowledgments
We thank Prof. Haibin Wang and colleagues at Xiamen University, Xiamen, China, for constructive insight and valuable advice. The authors expressed gratitude to all participants involved in this study.
Funding
The work was supported by National Key Research & Development Program of China (2016YFC1000202, 2018YFC1002804), Major Program of National Natural Science Foundation of China (81490743), General Program of National Natural Science Foundation of China (81671522, 81571406), Shandong Provincial Key Research and Development Program (2016GGH4522), and the Special Fund of Taishan Scholars Program of Shandong Province.
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All authors contributed to the study conception and design. Junhao Yan designed the study and finally approved the version to be submitted; Qian Zhang and Tianxiang Ni performed the study, analyzed the data, and wrote the paper; Yujie Dang performed the study; Lingling Ding and Jingjing Jiang collected samples and analyzed the data; Jing Li, Mingdi Xia, and Na Yu analyzed the data; and Jinlong Ma and Zi-Jiang Chen revised the paper. All authors read and approved the final manuscript.
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The study was approved by the Institutional Review Board of Center for Reproductive Medicine, Shandong University. Informed consent was obtained from all individual participants included in the study.
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The authors declare that they have no conflict of interest.
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Electronic supplementary material
Supplemental Table 1.
Primers for RT-qPCR (DOCX 16 kb)
Supplemental Fig 1.
Lentivirus-mediated miR-148a-3p overexpression or HOXC8 inhibition in HESCs (PNG 230 kb)
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Zhang, Q., Ni, T., Dang, Y. et al. MiR-148a-3p may contribute to flawed decidualization in recurrent implantation failure by modulating HOXC8. J Assist Reprod Genet 37, 2535–2544 (2020). https://doi.org/10.1007/s10815-020-01900-9
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DOI: https://doi.org/10.1007/s10815-020-01900-9