Studying the Oncosuppressive Functions of PTENP1 as a ceRNA.

Travis, G; Haddadi, N; Simpson, AM; Marsh, DJ; McGowan, EM; Nassif, NT

Studying the Oncosuppressive Functions of PTENP1 as a ceRNA.

Travis, G Haddadi, N Simpson, AM Marsh, DJ McGowan, EM Nassif, NT

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Publisher:: Springer US
Publication Type:: Journal Article
Citation:: Methods in molecular biology (Clifton, N.J.), 2021, 2324, pp. 165-185
Issue Date:: 2021-01

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Field	Value	Language
dc.contributor.author	Travis, G
dc.contributor.author	Haddadi, N
dc.contributor.author	Simpson, AM
dc.contributor.author	Marsh, DJ
dc.contributor.author	McGowan, EM
dc.contributor.author	Nassif, NT
dc.date.accessioned	2021-07-05T14:42:26Z
dc.date.available	2021-07-05T14:42:26Z
dc.date.issued	2021-01
dc.identifier.citation	Methods in molecular biology (Clifton, N.J.), 2021, 2324, pp. 165-185
dc.identifier.isbn	9781071615027
dc.identifier.issn	1064-3745
dc.identifier.issn	1940-6029
dc.identifier.uri	http://hdl.handle.net/10453/149833
dc.description.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<sup>®</sup> 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.
dc.format	Print
dc.language	eng
dc.publisher	Springer US
dc.relation	University of Technology SydneyUTSECRG04
dc.relation.ispartof	Methods in molecular biology (Clifton, N.J.)
dc.relation.isbasedon	10.1007/978-1-0716-1503-4_11
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0399 Other Chemical Sciences, 0601 Biochemistry and Cell Biology
dc.subject.classification	Developmental Biology
dc.title	Studying the Oncosuppressive Functions of PTENP1 as a ceRNA.
dc.type	Journal Article
utslib.citation.volume	2324
utslib.location.activity	United States
utslib.for	0399 Other Chemical Sciences
utslib.for	0601 Biochemistry and Cell Biology
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Centre for Health Technologies (CHT)
utslib.copyright.status	closed_access	*
dc.date.updated	2021-07-05T14:42:24Z
pubs.publication-status	Published
pubs.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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http://hdl.handle.net/10453/149833