Antisense oligonuleotides influences trophoblasts behaviors by changing LncNR_040117 expression in antiphospholipid antibody syndrome-induced recurrent pregnancy loss

Abstract

Objective

The primary objective of this study was to explore whether antisense oligonucleotides (ASOs) that reduce LncNR_040117 expression in patients with antiphospholipid antibody syndrome (APS)-induced recurrent pregnancy loss (RPL), and further decrease apoptosis and improve trophoblasts invasion through mitogen-activated protein kinase (MAPK) pathways. This paper aimed to provide a new strategy to treat APS-induced RPL.

Methods

In this study, we used quantitative reverse transcription-polymerase chain reaction (RT–qPCR) to analyze the expression level of LncNR 040117 in HTR-8/SVneo cells following transfection with ASOs. Then we utilized Western blotting to test the expression levels of interleukin-1β (IL-1β), intracellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and key molecules of MAPK pathways, including the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinases (JNK) and p38. In addition, we examined the HTR-8/SVneo cells apoptosis by cell apoptosis assay, and migration and invasion by transwell antibody assay. Each experiment was repeated three times. The data are presented as the means ± SDs, and statistical comparisons were performed using Student’s t-test. p < 0.05 was considered significant.

Result

Transfected with ASOs, LncNR_040117 was downregulated in trophoblasts compared with APS-induced RPL patients. And LncNR_040117 low expression induced IL-1β and downstream adhesion molecules ICAM-1 and VCAM-1expression level decreased, as well as MAPK pathways downregulation, including the ERK pathway, JNK pathway and p38/MAPK pathway. Furthermore, all these changes resulted in decreased apoptosis and increased migration and invasion of trophoblasts.

Conclusion

This study indicated that ASOs that decrease LncNR_040117 expression can reduce apoptosis and enhance the invasion and migration of trophoblasts by regulating the MAPK pathway.

Keywords:

• ASO
• LncNR_040117
• recurrent pregnancy loss
• antiphospholipid syndrome
• trophoblast cell

Acknowledgements

We thank Dr. Q Zhou for analysing platelet-derived microparticles (PMPs) in the peripheral blood of patients with APS-induced RPL by a lncRNA chip.

Authors’ contribution

X.T.W. and Q. Z. led the conception and design of the study. Q. Z. and D. W. performed the experiments. D. W. and Q. Z. was responsible for the analysis of the data. D. W. and Q. Z. wrote the paper. All authors have reviewed and approved the manuscript.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

Natural Science Foundation of Shandong Province (ZR2021QH197); National Natural Science Foundation of China (81571461 and 81873842); Key Research and Development Plan of Shandong Province (2018GSF118113).