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Potential in vitro therapeutic effects of targeting SP/NK1R system in cervical cancer

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Abstract

Background

Cervical cancer, an aggressive gynecological cancer, seriously threatens women’s health worldwide. It is recently reported that neuropeptide substance P (SP) regulates many tumor-associated processes through neurokinin-1 receptor (NK1R). Therefore, we used cervical cancer cell line (HeLa) to investigate the functional relevance of the SP/NK1R system in cervical cancer pathogenesis.

Methods

Cellular proliferation and cytotoxicity were analyzed by colorimetric MTT assay. Quantitative real-time PCR (qRT-PCR) was used to measure mRNA expression levels of desired genes. Cell cycle distribution and apoptosis were evaluated by flow cytometry. A wound-healing assay was employed to assess migration ability.

Results

We found that the truncated isoform of NK1R(NK1R-Tr) is the dominantly expressed form of the receptor in Hela cells. We also indicated that that SP increased HeLa cell proliferation while treatment with NK1R antagonist, aprepitant, inhibited HeLa cell viability in a dose and time-dependent manner. SP also alters the levels of cell cycle regulators (up-regulation of cyclin B1 along with downregulation of p21) and apoptosis-related genes (up-regulation of Bcl-2 along with downregulation of Bax) while aprepitant reversed these effects. Aprepitant also induced arrest within the G2 phase of the cell cycle and subsequent apoptosis. Furthermore, SP promoted the migrative phenotype of HeLa cells and increased MMP-2 and MMP-9 expression while aprepitant exposure significantly reversed these effects.

Conclusion

Collectively, our results indicate the importance of the SP / NK1R system in promoting both proliferative and migrative phenotypes of cervical cancer cells and suggest that aprepitant may be developed as a novel treatment for combating cervical cancer.

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Data availability

Data are available upon request.

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Funding

No official funding to declare. This research project was based on Mahtab Mozafari 's thesis and self-funded.

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Authors and Affiliations

  1. Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad, Iran

    Mahtab Mozafari & Reza Assaran Darban

  2. Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Safieh Ebrahimi & Seyed Isaac Hashemy

  3. Student Research Committee, Mashhad University of Medical Sciences, Mashhad, Iran

    Safieh Ebrahimi

  4. Surgical Oncology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Seyed Isaac Hashemy

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  1. Mahtab Mozafari
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  2. Safieh Ebrahimi
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  4. Seyed Isaac Hashemy
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Contributions

Conceptualization: SE, and SIH. Methodology: SE and MM. Formal analysis: SE, MM and RAD. Writing—original draft preparation: SE and MM.; writing—review and editing: All authors.; Supervision: SHA and RAD. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Reza Assaran Darban or Seyed Isaac Hashemy.

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Mozafari, M., Ebrahimi, S., Darban, R.A. et al. Potential in vitro therapeutic effects of targeting SP/NK1R system in cervical cancer. Mol Biol Rep 49, 1067–1076 (2022). https://doi.org/10.1007/s11033-021-06928-3

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