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The Effect of the cAMP Signaling Pathway on HTR8/SV-Neo Cell Line Proliferation, Invasion, and Migration After Treatment with Forskolin

  • Reproductive Biology: Original Article
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Abstract

Pre-eclampsia (PE) is thought to be related to placental dysfunction, particularly poor extravillous trophoblast (EVT) invasion and migration abilities. However, the pathogenic mechanism is not fully understood. This article describes the impact of the cyclic adenosine monophosphate(cAMP) signaling pathway on EVT behavior, focusing on EVT proliferation, invasion, and migration. Here, we used the HTR8/SV-neo cell line to study human EVT function in vitro. HTR8/SV-neo cells were treated with different concentrations of forskolin (cAMP pathway-specific agonist) to alter intracellular cAMP levels, and dimethyl sulfoxide (DMSO) was used as the control. First, a cAMP assay was performed to measure the cAMP concentration in HTR8/SV-neo cells treated with different forskolin concentrations, and cell proliferation was assessed by constructing cell growth curves and assessing colony formation. Cell invasion and migration were observed by Transwell experiments, and intracellular epithelial-mesenchymal transition (EMT) marker expression was evaluated by quantitative real-time polymerase chain reaction (qPCR) and Western blotting (WB). According to our research, the intracellular cAMP levels in HTR8/SV-neo cells were increased in a dose-dependent manner, and HTR8/SV-neo cell proliferation, invasion and migration were significantly enhanced. The expression of EMT and angiogenesis markers was upregulated. Additionally, with the increase in intracellular cAMP levels, the phosphorylation of intracellular mitogen-activated protein kinase (MAPK) signaling pathway components was significantly increased. These results suggested that the cAMP signaling pathway promoted the phosphorylation of MAPK signaling components, thus enhancing EVT functions, including proliferation, invasion, and migration, and to a certain extent, providing a novel direction for the treatment of PE patients.

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Data Availability

The data used to support the results of the study are included in the published paper.

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Acknowledgements

The authors sincerely thank all the participants in this study.

Funding

This study was financially supported by the Hainan Provincial Natural Science Foundation (No. ZDKJ2021037), the China Postdoctoral Science Foundation (No. 2021M691466) and the National Natural Science Foundation of China (No. 8220061871).

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Authors and Affiliations

  1. Hainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Key Laboratory of Reproductive Health Diseases Research and Translation (Hainan Medical University), Ministry of Education, The First Affiliated Hospital of Hainan Medical University, Hainan Medical University, 3 Longhua Road, Haikou, Hainan, 570102, China

    Chao Sun, Jiaoqi Mei, Hongyan Yi, Yanlin Ma & Yuanhua Huang

  2. Department of Reproductive Medicine, Hainan Provincial Clinical Research Center for Thalassemia, The First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou, Hainan, China

    Chao Sun, Jiaoqi Mei, Hongyan Yi, Yanlin Ma & Yuanhua Huang

  3. National Center for International Research “China-Myanmar Joint Research Center for Prevention and Treatment of Regional Major Disease” by the Ministry of Science and Technology of China, The First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou, Hainan, China

    Chao Sun, Jiaoqi Mei, Hongyan Yi, Yanlin Ma & Yuanhua Huang

  4. Haikou Key Laboratory for Preservation of Human Genetic Resource, The First Affiliated Hospital of Hainan Medical University, Hainan Medical University, Haikou, Hainan, China

    Chao Sun, Jiaoqi Mei, Hongyan Yi, Yanlin Ma & Yuanhua Huang

  5. Center for Reproductive Medicine, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China

    Mengyi Song

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  1. Chao Sun
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Contributions

Yuanhua Huang and Yanlin Ma were responsible for the conception and design of the study and the experiments, while data collection was performed by Jiaoqi Mei and Mengyi Song. Yanhong Yi and ChaoSun designed the experiments and drafted the manuscript. The final version of the manuscript was approved by all the authors.

Corresponding authors

Correspondence to Yanlin Ma or Yuanhua Huang.

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Ethics Approval

All the studies were approved by the Medical Ethics Committee of First Affiliated Hospital of Hainan Medical University(2022-KYL-039), and performed in accordance with the Declaration of Helsinki.

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Chao Sun and Jiaoqi Mei share first authorship

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Sun, C., Mei, J., Yi, H. et al. The Effect of the cAMP Signaling Pathway on HTR8/SV-Neo Cell Line Proliferation, Invasion, and Migration After Treatment with Forskolin. Reprod. Sci. (2023). https://doi.org/10.1007/s43032-023-01396-5

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  • DOI: https://doi.org/10.1007/s43032-023-01396-5

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