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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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Evaluation of the CSC Features of the MDA-MB-231 D3H2LN and C3LND Cell Lines(A) Mammosphere quantification in the modestly metastatic, parental MDA-MB-231 D3H2LN and the derived, highly metastatic C3LND cell lines (n = 3 independent experiments).(B) The qRT-PCR analysis of Δ133p53 isoform expression in MDA-MB-231 D3H2LN and C3LND cells (n = 4 independent experiments).(C) The qRT-PCR quantification of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 D3H2LN and C3LND cells (n = 4 independent experiments).(D) Western blot analysis of Δ133p53β-Flag transduced in MDA-MB-231 D3H2LN cells (Flag antibody).(E) The qRT-PCR analysis of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 D3H2LN cells after Δ133p53β overexpression (n = 4 independent experiments).(F) Mammosphere quantification in MDA-MB-231 D3H2LN cells that overexpress Δ133p53β (n = 3 independent experiments).(G) Mammosphere quantification in MDA-MB-231 C3LND transduced with Sh3 (n = 3 independent experiments).(H) The qRT-PCR analysis of Δ133p53 isoform expression in MDA-MB-231 C3LND transduced with Sh3 (n = 3 independent experiments).(I) The qRT-PCR quantification of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 C3LND cells transduced with Sh3 (n = 4 independent experiments).(J) Representative FACS dot plots for the double labeling of CD44 and CD24 in MDA-MB-231 C3LND transduced with Sh Luc (Control) or Sh3 (n = 3 independent experiments).(K) Bioluminescence ventral images of mice injected with MDA-MB-231 C3LND cells transduced with Sh3 or control. Pseudocolor scale bars show relative changes at metastatic sites.(L) Quantification of distant metastasis in brain and femur using bioluminescence imaging (n = 7/5) 25 days after the implantation. Bars represent mean ± SEM of biological replicates.
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fig3: Evaluation of the CSC Features of the MDA-MB-231 D3H2LN and C3LND Cell Lines(A) Mammosphere quantification in the modestly metastatic, parental MDA-MB-231 D3H2LN and the derived, highly metastatic C3LND cell lines (n = 3 independent experiments).(B) The qRT-PCR analysis of Δ133p53 isoform expression in MDA-MB-231 D3H2LN and C3LND cells (n = 4 independent experiments).(C) The qRT-PCR quantification of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 D3H2LN and C3LND cells (n = 4 independent experiments).(D) Western blot analysis of Δ133p53β-Flag transduced in MDA-MB-231 D3H2LN cells (Flag antibody).(E) The qRT-PCR analysis of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 D3H2LN cells after Δ133p53β overexpression (n = 4 independent experiments).(F) Mammosphere quantification in MDA-MB-231 D3H2LN cells that overexpress Δ133p53β (n = 3 independent experiments).(G) Mammosphere quantification in MDA-MB-231 C3LND transduced with Sh3 (n = 3 independent experiments).(H) The qRT-PCR analysis of Δ133p53 isoform expression in MDA-MB-231 C3LND transduced with Sh3 (n = 3 independent experiments).(I) The qRT-PCR quantification of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 C3LND cells transduced with Sh3 (n = 4 independent experiments).(J) Representative FACS dot plots for the double labeling of CD44 and CD24 in MDA-MB-231 C3LND transduced with Sh Luc (Control) or Sh3 (n = 3 independent experiments).(K) Bioluminescence ventral images of mice injected with MDA-MB-231 C3LND cells transduced with Sh3 or control. Pseudocolor scale bars show relative changes at metastatic sites.(L) Quantification of distant metastasis in brain and femur using bioluminescence imaging (n = 7/5) 25 days after the implantation. Bars represent mean ± SEM of biological replicates.

Mentions: Evaluation of mammosphere formation in D3H2LN and C3LND cells showed that C3LND cells formed two times more mammospheres (Figure 3A). Similarly, Δ133p53 isoform expression was 3-fold higher and OCT3/4, NANOG, and SOX2 levels were 2- to 3-fold higher in C3LND (Figures 3B and 3C). C-MYC expression was comparable in the two cells lines. We then asked whether pluripotency factor expression could be affected by changes in Δ133p53 expression. Overexpression of Δ133p53β in D3H2LN cells resulted in a significant increase of OCT3/4, NANOG, and SOX2 expression, whereas C-MYC level was not affected, consistent with data in MCF-7 cells (Figures 3D and 3E). Similar results were obtained in C3LND cells (Figure S3F). In complete agreement with observations in MCF-7, Δ133p53β overexpression in D3H2LN cells resulted in a significant increase of mammosphere formation (Figure 3F).


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)

Evaluation of the CSC Features of the MDA-MB-231 D3H2LN and C3LND Cell Lines(A) Mammosphere quantification in the modestly metastatic, parental MDA-MB-231 D3H2LN and the derived, highly metastatic C3LND cell lines (n = 3 independent experiments).(B) The qRT-PCR analysis of Δ133p53 isoform expression in MDA-MB-231 D3H2LN and C3LND cells (n = 4 independent experiments).(C) The qRT-PCR quantification of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 D3H2LN and C3LND cells (n = 4 independent experiments).(D) Western blot analysis of Δ133p53β-Flag transduced in MDA-MB-231 D3H2LN cells (Flag antibody).(E) The qRT-PCR analysis of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 D3H2LN cells after Δ133p53β overexpression (n = 4 independent experiments).(F) Mammosphere quantification in MDA-MB-231 D3H2LN cells that overexpress Δ133p53β (n = 3 independent experiments).(G) Mammosphere quantification in MDA-MB-231 C3LND transduced with Sh3 (n = 3 independent experiments).(H) The qRT-PCR analysis of Δ133p53 isoform expression in MDA-MB-231 C3LND transduced with Sh3 (n = 3 independent experiments).(I) The qRT-PCR quantification of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 C3LND cells transduced with Sh3 (n = 4 independent experiments).(J) Representative FACS dot plots for the double labeling of CD44 and CD24 in MDA-MB-231 C3LND transduced with Sh Luc (Control) or Sh3 (n = 3 independent experiments).(K) Bioluminescence ventral images of mice injected with MDA-MB-231 C3LND cells transduced with Sh3 or control. Pseudocolor scale bars show relative changes at metastatic sites.(L) Quantification of distant metastasis in brain and femur using bioluminescence imaging (n = 7/5) 25 days after the implantation. Bars represent mean ± SEM of biological replicates.
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fig3: Evaluation of the CSC Features of the MDA-MB-231 D3H2LN and C3LND Cell Lines(A) Mammosphere quantification in the modestly metastatic, parental MDA-MB-231 D3H2LN and the derived, highly metastatic C3LND cell lines (n = 3 independent experiments).(B) The qRT-PCR analysis of Δ133p53 isoform expression in MDA-MB-231 D3H2LN and C3LND cells (n = 4 independent experiments).(C) The qRT-PCR quantification of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 D3H2LN and C3LND cells (n = 4 independent experiments).(D) Western blot analysis of Δ133p53β-Flag transduced in MDA-MB-231 D3H2LN cells (Flag antibody).(E) The qRT-PCR analysis of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 D3H2LN cells after Δ133p53β overexpression (n = 4 independent experiments).(F) Mammosphere quantification in MDA-MB-231 D3H2LN cells that overexpress Δ133p53β (n = 3 independent experiments).(G) Mammosphere quantification in MDA-MB-231 C3LND transduced with Sh3 (n = 3 independent experiments).(H) The qRT-PCR analysis of Δ133p53 isoform expression in MDA-MB-231 C3LND transduced with Sh3 (n = 3 independent experiments).(I) The qRT-PCR quantification of C-MYC, OCT3/4, NANOG, and SOX2 expression in MDA-MB-231 C3LND cells transduced with Sh3 (n = 4 independent experiments).(J) Representative FACS dot plots for the double labeling of CD44 and CD24 in MDA-MB-231 C3LND transduced with Sh Luc (Control) or Sh3 (n = 3 independent experiments).(K) Bioluminescence ventral images of mice injected with MDA-MB-231 C3LND cells transduced with Sh3 or control. Pseudocolor scale bars show relative changes at metastatic sites.(L) Quantification of distant metastasis in brain and femur using bioluminescence imaging (n = 7/5) 25 days after the implantation. Bars represent mean ± SEM of biological replicates.
Mentions: Evaluation of mammosphere formation in D3H2LN and C3LND cells showed that C3LND cells formed two times more mammospheres (Figure 3A). Similarly, Δ133p53 isoform expression was 3-fold higher and OCT3/4, NANOG, and SOX2 levels were 2- to 3-fold higher in C3LND (Figures 3B and 3C). C-MYC expression was comparable in the two cells lines. We then asked whether pluripotency factor expression could be affected by changes in Δ133p53 expression. Overexpression of Δ133p53β in D3H2LN cells resulted in a significant increase of OCT3/4, NANOG, and SOX2 expression, whereas C-MYC level was not affected, consistent with data in MCF-7 cells (Figures 3D and 3E). Similar results were obtained in C3LND cells (Figure S3F). In complete agreement with observations in MCF-7, Δ133p53β overexpression in D3H2LN cells resulted in a significant increase of mammosphere formation (Figure 3F).

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