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The p53 isoform Δ133p53β promotes cancer stem cell potential.

Arsic N, Gadea G, Lagerqvist EL, Busson M, Cahuzac N, Brock C, Hollande F, Gire V, Pannequin J, Roux P - Stem Cell Reports (2015)

Bottom Line: Here we report that Δ133p53β, a TP53 splice variant, enhanced cancer cell stemness in MCF-7 breast cancer cells, while its depletion reduced it.Like in MCF-7 cells, SOX2, OCT3/4, and NANOG expression were positively regulated by Δ133p53β in these cells.Our findings show that Δ133p53β supports CSC potential.

View Article: PubMed Central - PubMed

Affiliation: Centre National de la Recherche Scientifique, UMR 5237, Centre de Recherche en Biochimie Macromoléculaire, Université Montpellier, 1919 route de Mende, 34293 Montpellier Cedex 5, France.

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The Isoform Δ133p53β Promotes CSC Potential in MCF-7 Cells(A and B) Mammosphere quantification in MCF-7 cells after silencing with Sh2 (shRNAs against the TA and Δ40 isoforms) or with Sh2 and Sh6 (against the 3′ end of the α isoforms) (A) and western blot analysis to confirm p53 depletion in the corresponding cell cultures (B) (n = 3 independent experiments).(C) Representative fluorescence-activated cell sorting (FACS) dot blots for the double labeling of CD24 and CD44 in MCF-7 transduced with Sh Luc (Control), Sh2, or Sh2 + 6.(D and E) Mammosphere quantification in MCF-7 cells after Δ133p53β or γ overexpression (D) and qRT-PCR analysis of C-MYC, SOX2, OCT3/4, and NANOG (E) expression in the corresponding cells (n = 4 independent experiments).(F) Mammosphere quantification in MCF-7 cells that overexpress Δ133p53β after harvesting and re-plating of the primary mammospheres (n = 4 independent experiments).(G) Mammosphere quantification in MCF-7 cells in which all p53 isoforms have been silenced with Sh1 and after expression in the same cells of Sh1-resistant Δ133p53β (n = 3 independent experiments).
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fig2: The Isoform Δ133p53β Promotes CSC Potential in MCF-7 Cells(A and B) Mammosphere quantification in MCF-7 cells after silencing with Sh2 (shRNAs against the TA and Δ40 isoforms) or with Sh2 and Sh6 (against the 3′ end of the α isoforms) (A) and western blot analysis to confirm p53 depletion in the corresponding cell cultures (B) (n = 3 independent experiments).(C) Representative fluorescence-activated cell sorting (FACS) dot blots for the double labeling of CD24 and CD44 in MCF-7 transduced with Sh Luc (Control), Sh2, or Sh2 + 6.(D and E) Mammosphere quantification in MCF-7 cells after Δ133p53β or γ overexpression (D) and qRT-PCR analysis of C-MYC, SOX2, OCT3/4, and NANOG (E) expression in the corresponding cells (n = 4 independent experiments).(F) Mammosphere quantification in MCF-7 cells that overexpress Δ133p53β after harvesting and re-plating of the primary mammospheres (n = 4 independent experiments).(G) Mammosphere quantification in MCF-7 cells in which all p53 isoforms have been silenced with Sh1 and after expression in the same cells of Sh1-resistant Δ133p53β (n = 3 independent experiments).

Mentions: Indeed, mammosphere formation was significantly increased in MCF-7 cells that expressed only the Δ133p53β and Δ133p53γ isoforms following concomitant transduction with Sh2 and Sh6 (Figures 2A and 2B). To confirm that sphere increase was indicative of the CSC phenotype, we analyzed the proportion of CD44+/CD24− cells, because this subpopulation of cancer cells are considered to have CSC properties. Similar to mammosphere formation variations, the proportion of CD44+/CD24− cells was not affected by TAp53 and Δ40p53 isoform silencing with Sh2, whereas it was increased by co-transduction of Sh2 and Sh6 (Figure 2C).


The p53 isoform Δ133p53β promotes cancer stem cell potential.

Arsic N, Gadea G, Lagerqvist EL, Busson M, Cahuzac N, Brock C, Hollande F, Gire V, Pannequin J, Roux P - Stem Cell Reports (2015)

The Isoform Δ133p53β Promotes CSC Potential in MCF-7 Cells(A and B) Mammosphere quantification in MCF-7 cells after silencing with Sh2 (shRNAs against the TA and Δ40 isoforms) or with Sh2 and Sh6 (against the 3′ end of the α isoforms) (A) and western blot analysis to confirm p53 depletion in the corresponding cell cultures (B) (n = 3 independent experiments).(C) Representative fluorescence-activated cell sorting (FACS) dot blots for the double labeling of CD24 and CD44 in MCF-7 transduced with Sh Luc (Control), Sh2, or Sh2 + 6.(D and E) Mammosphere quantification in MCF-7 cells after Δ133p53β or γ overexpression (D) and qRT-PCR analysis of C-MYC, SOX2, OCT3/4, and NANOG (E) expression in the corresponding cells (n = 4 independent experiments).(F) Mammosphere quantification in MCF-7 cells that overexpress Δ133p53β after harvesting and re-plating of the primary mammospheres (n = 4 independent experiments).(G) Mammosphere quantification in MCF-7 cells in which all p53 isoforms have been silenced with Sh1 and after expression in the same cells of Sh1-resistant Δ133p53β (n = 3 independent experiments).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400643&req=5

fig2: The Isoform Δ133p53β Promotes CSC Potential in MCF-7 Cells(A and B) Mammosphere quantification in MCF-7 cells after silencing with Sh2 (shRNAs against the TA and Δ40 isoforms) or with Sh2 and Sh6 (against the 3′ end of the α isoforms) (A) and western blot analysis to confirm p53 depletion in the corresponding cell cultures (B) (n = 3 independent experiments).(C) Representative fluorescence-activated cell sorting (FACS) dot blots for the double labeling of CD24 and CD44 in MCF-7 transduced with Sh Luc (Control), Sh2, or Sh2 + 6.(D and E) Mammosphere quantification in MCF-7 cells after Δ133p53β or γ overexpression (D) and qRT-PCR analysis of C-MYC, SOX2, OCT3/4, and NANOG (E) expression in the corresponding cells (n = 4 independent experiments).(F) Mammosphere quantification in MCF-7 cells that overexpress Δ133p53β after harvesting and re-plating of the primary mammospheres (n = 4 independent experiments).(G) Mammosphere quantification in MCF-7 cells in which all p53 isoforms have been silenced with Sh1 and after expression in the same cells of Sh1-resistant Δ133p53β (n = 3 independent experiments).
Mentions: Indeed, mammosphere formation was significantly increased in MCF-7 cells that expressed only the Δ133p53β and Δ133p53γ isoforms following concomitant transduction with Sh2 and Sh6 (Figures 2A and 2B). To confirm that sphere increase was indicative of the CSC phenotype, we analyzed the proportion of CD44+/CD24− cells, because this subpopulation of cancer cells are considered to have CSC properties. Similar to mammosphere formation variations, the proportion of CD44+/CD24− cells was not affected by TAp53 and Δ40p53 isoform silencing with Sh2, whereas it was increased by co-transduction of Sh2 and Sh6 (Figure 2C).

Bottom Line: Here we report that Δ133p53β, a TP53 splice variant, enhanced cancer cell stemness in MCF-7 breast cancer cells, while its depletion reduced it.Like in MCF-7 cells, SOX2, OCT3/4, and NANOG expression were positively regulated by Δ133p53β in these cells.Our findings show that Δ133p53β supports CSC potential.

View Article: PubMed Central - PubMed

Affiliation: Centre National de la Recherche Scientifique, UMR 5237, Centre de Recherche en Biochimie Macromoléculaire, Université Montpellier, 1919 route de Mende, 34293 Montpellier Cedex 5, France.

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Related in: MedlinePlus