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Studying the Oncosuppressive Functions of PTENP1 as a ceRNA

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Pseudogenes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2324))

Abstract

PTENP1 is a processed pseudogene of the tumour suppressor phosphatase and tensin homolog deleted on chromosome 10 (PTEN). It functions posttranscriptionally to regulate PTEN by acting as a sponge for microRNAs that target PTEN. PTENP1 therefore functions as a competitive endogenous RNA (ceRNA), competing with PTEN for binding of microRNAs (miRNA) and thereby modulating PTEN cellular abundance. Studies of the overexpression of PTENP1 all confirm its oncosuppressive function to be mediated through the suppression of cell proliferation, induction of apoptosis, and inhibition of cell migration and invasion of cancer cells of differing types. These oncosuppressive functions are a direct consequence of miRNA binding by PTENP1 and the subsequent liberation of PTEN from miRNA induced suppression. In this chapter, we will focus initially on the description of a high efficiency transient transfection method to introduce and overexpress PTENP1 in the cell type of interest, followed by accurate methodologies to measure transfection efficiency by flow cytometry. We will then continue to describe two methods to analyze cell proliferation, namely the CCK-8 assay and Click-iT® EdU assay. Due to commonalities in the manifestation of the oncosuppressive effects of PTENP1, mediated through its role as a ceRNA, the methods presented in this chapter will have wide applicability to a variety of different cell types.

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Acknowledgments

Glena Travis is supported by the Australian Research Training Program Stipend Scholarship, and Nahal Haddadi was supported by a University of Technology Sydney International Research Scholarship.

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Authors and Affiliations

  1. School of Life Sciences, Faculty of Science, University of Technology Sydney, Ultimo, NSW, Australia

    Glena Travis, Nahal Haddadi, Ann M. Simpson, Deborah J. Marsh, Eileen M. McGowan & Najah T. Nassif

  2. Centre for Health Technologies, University of Technology Sydney, Ultimo, NSW, Australia

    Ann M. Simpson, Deborah J. Marsh & Najah T. Nassif

  3. Northern Clinical School, Kolling Institute, Faculty of Medicine and Health, University of Sydney, Sydney, NSW, Australia

    Deborah J. Marsh

Authors
  1. Glena Travis
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  2. Nahal Haddadi
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  3. Ann M. Simpson
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  4. Deborah J. Marsh
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  5. Eileen M. McGowan
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  6. Najah T. Nassif
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Corresponding author

Correspondence to Najah T. Nassif .

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Editors and Affiliations

  1. Oncogenomics Unit, Core Research Laboratory, ISPRO, Institute of Clinical Physiology (IFC), CNR, Pisa, Italy

    Laura Poliseno

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Travis, G., Haddadi, N., Simpson, A.M., Marsh, D.J., McGowan, E.M., Nassif, N.T. (2021). Studying the Oncosuppressive Functions of PTENP1 as a ceRNA . In: Poliseno, L. (eds) Pseudogenes. Methods in Molecular Biology, vol 2324. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1503-4_11

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  • DOI: https://doi.org/10.1007/978-1-0716-1503-4_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1502-7

  • Online ISBN: 978-1-0716-1503-4

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