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miR-195-5p Targets CDK1 To Regulate New DNA Synthesis and Inhibit the Proliferation of Hepatocellular Carcinoma Cells

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Abstract

In cell biological functions and viability, cyclin-dependent kinase 1 (CDK1) takes an essential part. miR-195-5p is pivotal in pathogenesis and development of hepatocellular carcinoma (HCC). But in HCC, whether there is a connection between CDK1 and miR-195-5p remains an unanswered question. In view of this, this study focuses on exploring the mechanism of miR-195-5p/CDK1 in the progression of HCC. The bioinformatics method was applied to predict target mRNA and upstream miRNAs, and further analyzes the signal enrichment pathway of target mRNA. We utilized qRT-PCR and Western blot for detecting expression of genes, as well as their corresponding protein levels. Cell cycle was assayed through flow cytometry. As for the examination of DNA replication, the EDU staining was employed. Cell proliferation was determined via plate colony formation assay. The combined application of bioinformatics analysis and dual-luciferase gene assay assisted in figuring out the binding relationship between miR-195-5p and CDK1. DNA damage was marked by immunofluorescence staining. CDK1 was overexpressed in HCC cells, and enriched in cell cycle and DNA replication pathway. Silencing CDK1 modulated cell cycle of HCC cells and inhibited DNA replication and proliferation. In HCC cells, miR-195-5p targeted and reduced CDK1 expression, inhibited the G1 phase-to-S phase transition, induced DNA damage response, and inhibited DNA replication and proliferation. miR-195-5p targeted CDK1 and repressed synthesis of new DNA in HCC cells, thus restraining HCC cell proliferation.

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Data Availability

The datasets generated and analyzed during the current study are not publicly available but are available from the corresponding author on reasonable request.

References

  1. Pinero, F., Dirchwolf, M., & Pessoa, M. G. (2020). Biomarkers in hepatocellular carcinoma: Diagnosis, prognosis and treatment response assessment. Cells, 9, 1370. https://doi.org/10.3390/cells9061370

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Sim, H. W., & Knox, J. (2018). Hepatocellular carcinoma in the era of immunotherapy. Current Problems in Cancer, 42, 40–48. https://doi.org/10.1016/j.currproblcancer.2017.10.007

    Article  PubMed  Google Scholar 

  3. Santamaria, D., et al. (2007). Cdk1 is sufficient to drive the mammalian cell cycle. Nature, 448, 811–815. https://doi.org/10.1038/nature06046

    Article  CAS  PubMed  Google Scholar 

  4. Garnier, D., Loyer, P., Ribault, C., Guguen-Guillouzo, C., & Corlu, A. (2009). Cyclin-dependent kinase 1 plays a critical role in DNA replication control during rat liver regeneration. Hepatology, 50, 1946–1956. https://doi.org/10.1002/hep.23225

    Article  CAS  PubMed  Google Scholar 

  5. Johnson, N., et al. (2009). Cdk1 participates in BRCA1-dependent S phase checkpoint control in response to DNA damage. Molecular Cell, 35, 327–339. https://doi.org/10.1016/j.molcel.2009.06.036

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Chow, J. P., Poon, R. Y., & Ma, H. T. (2011). Inhibitory phosphorylation of cyclin-dependent kinase 1 as a compensatory mechanism for mitosis exit. Molecular and Cellular Biology, 31, 1478–1491. https://doi.org/10.1128/MCB.00891-10

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Sung, W. W., et al. (2014). High nuclear/cytoplasmic ratio of Cdk1 expression predicts poor prognosis in colorectal cancer patients. BMC Cancer, 14, 951. https://doi.org/10.1186/1471-2407-14-951

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Willder, J. M., et al. (2013). Androgen receptor phosphorylation at serine 515 by Cdk1 predicts biochemical relapse in prostate cancer patients. British Journal of Cancer, 108, 139–148. https://doi.org/10.1038/bjc.2012.480

    Article  CAS  PubMed  Google Scholar 

  9. Danhier, F., Ucakar, B., Magotteaux, N., Brewster, M. E., & Preat, V. (2010). Active and passive tumor targeting of a novel poorly soluble cyclin dependent kinase inhibitor, JNJ-7706621. International Journal of Pharmaceutics, 392, 20–28. https://doi.org/10.1016/j.ijpharm.2010.03.018

    Article  CAS  PubMed  Google Scholar 

  10. Jin, J., et al. (2020). LINC00346 Acts as a competing endogenous Rna regulating development of hepatocellular carcinoma via modulating CDK1/CCNB1 AXIS. Front Bioeng Biotechnol, 8, 54. https://doi.org/10.3389/fbioe.2020.00054

    Article  PubMed  PubMed Central  Google Scholar 

  11. Xu, T., et al. (2009). MicroRNA-195 suppresses tumorigenicity and regulates G1/S transition of human hepatocellular carcinoma cells. Hepatology, 50, 113–121. https://doi.org/10.1002/hep.22919

    Article  CAS  PubMed  Google Scholar 

  12. Ozata, D. M., et al. (2011). The role of microRNA deregulation in the pathogenesis of adrenocortical carcinoma. Endocrine-Related Cancer, 18, 643–655. https://doi.org/10.1530/ERC-11-0082

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Zhang, Q. Q., et al. (2012). MicroRNA-195 plays a tumor-suppressor role in human glioblastoma cells by targeting signaling pathways involved in cellular proliferation and invasion. Neuro-Oncology, 14, 278–287. https://doi.org/10.1093/neuonc/nor216

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Li, D., et al. (2011). Analysis of MiR-195 and MiR-497 expression, regulation and role in breast cancer. Clinical Cancer Research, 17, 1722–1730. https://doi.org/10.1158/1078-0432.CCR-10-1800

    Article  CAS  PubMed  Google Scholar 

  15. Huang, D. P., et al. (2021). Bioinformatics analyses of potential miRNA-mRNA regulatory axis in HBV-related hepatocellular carcinoma. International Journal of Medical Sciences, 18, 335–346. https://doi.org/10.7150/ijms.50126

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Yang, W., et al. (2016). Accumulation of cytoplasmic Cdk1 is associated with cancer growth and survival rate in epithelial ovarian cancer. Oncotarget, 7, 49481–49497. https://doi.org/10.18632/oncotarget.10373

    Article  PubMed  PubMed Central  Google Scholar 

  17. Piao, J., et al. (2019). High expression of CDK1 and BUB1 predicts poor prognosis of pancreatic ductal adenocarcinoma. Gene, 701, 15–22. https://doi.org/10.1016/j.gene.2019.02.081

    Article  CAS  PubMed  Google Scholar 

  18. Wu, C. X., et al. (2018). Blocking CDK1/PDK1/beta-catenin signaling by CDK1 inhibitor RO3306 increased the efficacy of sorafenib treatment by targeting cancer stem cells in a preclinical model of hepatocellular carcinoma. Theranostics, 8, 3737–3750. https://doi.org/10.7150/thno.25487

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Xu, H., et al. (2015). MicroRNA-195-5p acts as an anti-oncogene by targeting PHF19 in hepatocellular carcinoma. Oncology Reports, 34, 175–182. https://doi.org/10.3892/or.2015.3957

    Article  CAS  PubMed  Google Scholar 

  20. Johnson, N., et al. (2011). Compromised CDK1 activity sensitizes BRCA-proficient cancers to PARP inhibition. Nature Medicine, 17, 875–882. https://doi.org/10.1038/nm.2377

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Radiological Intervention Center, Department of Radiology, Xiangya Hospital, Central South University, 87 Xiangya Road, Kai Fu District Changsha City, Hunan Province, 410008, China

    Chunhui Zhou & Haiping Li

  2. The Tumor Hospital of SUMC, Cancer Hospital of Shantou University Medical College, Shantou City, Guangdong Province, 515000, China

    Sujuan Zhu

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  1. Chunhui Zhou
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  2. Sujuan Zhu
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Contributions

Chunhui Zhou: conceptualization, methodology, software, data curation, writing- original draft preparation.

Sujuan Zhu: visualization, investigation, supervision, validation.

Haiping Li: writing- reviewing and editing.

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Correspondence to Haiping Li.

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Zhou, C., Zhu, S. & Li, H. miR-195-5p Targets CDK1 To Regulate New DNA Synthesis and Inhibit the Proliferation of Hepatocellular Carcinoma Cells. Appl Biochem Biotechnol 195, 3477–3490 (2023). https://doi.org/10.1007/s12010-022-04279-8

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