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Wig-1 regulates cell cycle arrest and cell death through the p53 targets FAS and 14-3-3σ.

Bersani C, Xu LD, Vilborg A, Lui WO, Wiman KG - Oncogene (2014)

Bottom Line: We identified 2447 transcripts with >fourfold differential expression between Wig-1 and control small interfering (si)RNA-treated HCT116 cells.We found that Wig-1 regulates FAS and 14-3-3σ mRNA independently of p53.We show that Wig-1 binds to FAS mRNA 3'-UTR and decreases its stability through an ARE in the 3'-UTR.

View Article: PubMed Central - PubMed

Affiliation: Cancer Center Karolinska (CCK), Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden.

ABSTRACT
Wig-1, also known as ZMAT3, is a p53 target gene that encodes an RNA-binding zinc-finger protein involved in the regulation of mRNA stability through binding to AU-rich elements (AREs). We have used microarray analysis to identify novel Wig-1 target mRNAs. We identified 2447 transcripts with >fourfold differential expression between Wig-1 and control small interfering (si)RNA-treated HCT116 cells. Several p53 target genes were among the deregulated transcripts. We found that Wig-1 regulates FAS and 14-3-3σ mRNA independently of p53. We show that Wig-1 binds to FAS mRNA 3'-UTR and decreases its stability through an ARE in the 3'-UTR. Depletion of Wig-1 was associated with increased cell death and reduced cell cycle arrest upon DNA damage. Our results suggest a role of Wig-1 as a survival factor that directs the p53 stress response toward cell cycle arrest rather than apoptosis through the regulation of FAS and 14-3-3σ mRNA levels.

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Wig-1 knockdown leads to cell death rather than cell cycle arrest upon cellular stress in HCT116 cells. FACS analysis of cell cycle distribution in HCT116 p53+/+ and p53−/− cells transfected with control siRNA or siRNA targeting Wig-1 (siW1) demonstrates a decrease in the G2 (a) and an increase in the sub-G1 population (b) after treatment with cisplatin (5 μM 24 h). Annexin V staining confirmed that the observed cell death was due to apoptosis (c). Decreased proliferation/cell viability 4 days after Wig-1 knockdown and 5 μM cisplatin treatment or no treatment is shown by live cell number count using Trypan blue exclusion (d) and Giemsa staining of fixed cells in cell culture dishes (e). Columns and error bars represent the mean±s.d.; n=3; ***P<0.001; **P<0.01.
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fig2: Wig-1 knockdown leads to cell death rather than cell cycle arrest upon cellular stress in HCT116 cells. FACS analysis of cell cycle distribution in HCT116 p53+/+ and p53−/− cells transfected with control siRNA or siRNA targeting Wig-1 (siW1) demonstrates a decrease in the G2 (a) and an increase in the sub-G1 population (b) after treatment with cisplatin (5 μM 24 h). Annexin V staining confirmed that the observed cell death was due to apoptosis (c). Decreased proliferation/cell viability 4 days after Wig-1 knockdown and 5 μM cisplatin treatment or no treatment is shown by live cell number count using Trypan blue exclusion (d) and Giemsa staining of fixed cells in cell culture dishes (e). Columns and error bars represent the mean±s.d.; n=3; ***P<0.001; **P<0.01.

Mentions: As the absence of Wig-1 caused an increase in the proapoptotic factor FAS and a decrease in the cell cycle arrest inducer 14-3-3σ, we hypothesized that Wig-1 might have a role in controlling the outcome of cellular stress. To investigate this possibility, we silenced Wig-1 in HCT116 p53+/+ and p53−/− cells and analyzed the effect on both cell cycle distribution and cell death by fluorescence activated cell sorting. We used cisplatin as a stress-inducing agent, as it induced G2 cell cycle arrest but no cell death. Upon cisplatin-induced stress, Wig-1 silencing led to a decrease in the G2 fraction of HCT116 p53+/+ (14%) and HCT116 p53−/− (15%) (P-value=0.001) cells relative to the control-treated cells (Figure 2a). Moreover, Wig-1 knockdown resulted in a significant increase in the sub-G1 population in both HCT116 p53+/+ and p53−/− cells (Figure 2b).


Wig-1 regulates cell cycle arrest and cell death through the p53 targets FAS and 14-3-3σ.

Bersani C, Xu LD, Vilborg A, Lui WO, Wiman KG - Oncogene (2014)

Wig-1 knockdown leads to cell death rather than cell cycle arrest upon cellular stress in HCT116 cells. FACS analysis of cell cycle distribution in HCT116 p53+/+ and p53−/− cells transfected with control siRNA or siRNA targeting Wig-1 (siW1) demonstrates a decrease in the G2 (a) and an increase in the sub-G1 population (b) after treatment with cisplatin (5 μM 24 h). Annexin V staining confirmed that the observed cell death was due to apoptosis (c). Decreased proliferation/cell viability 4 days after Wig-1 knockdown and 5 μM cisplatin treatment or no treatment is shown by live cell number count using Trypan blue exclusion (d) and Giemsa staining of fixed cells in cell culture dishes (e). Columns and error bars represent the mean±s.d.; n=3; ***P<0.001; **P<0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150987&req=5

fig2: Wig-1 knockdown leads to cell death rather than cell cycle arrest upon cellular stress in HCT116 cells. FACS analysis of cell cycle distribution in HCT116 p53+/+ and p53−/− cells transfected with control siRNA or siRNA targeting Wig-1 (siW1) demonstrates a decrease in the G2 (a) and an increase in the sub-G1 population (b) after treatment with cisplatin (5 μM 24 h). Annexin V staining confirmed that the observed cell death was due to apoptosis (c). Decreased proliferation/cell viability 4 days after Wig-1 knockdown and 5 μM cisplatin treatment or no treatment is shown by live cell number count using Trypan blue exclusion (d) and Giemsa staining of fixed cells in cell culture dishes (e). Columns and error bars represent the mean±s.d.; n=3; ***P<0.001; **P<0.01.
Mentions: As the absence of Wig-1 caused an increase in the proapoptotic factor FAS and a decrease in the cell cycle arrest inducer 14-3-3σ, we hypothesized that Wig-1 might have a role in controlling the outcome of cellular stress. To investigate this possibility, we silenced Wig-1 in HCT116 p53+/+ and p53−/− cells and analyzed the effect on both cell cycle distribution and cell death by fluorescence activated cell sorting. We used cisplatin as a stress-inducing agent, as it induced G2 cell cycle arrest but no cell death. Upon cisplatin-induced stress, Wig-1 silencing led to a decrease in the G2 fraction of HCT116 p53+/+ (14%) and HCT116 p53−/− (15%) (P-value=0.001) cells relative to the control-treated cells (Figure 2a). Moreover, Wig-1 knockdown resulted in a significant increase in the sub-G1 population in both HCT116 p53+/+ and p53−/− cells (Figure 2b).

Bottom Line: We identified 2447 transcripts with >fourfold differential expression between Wig-1 and control small interfering (si)RNA-treated HCT116 cells.We found that Wig-1 regulates FAS and 14-3-3σ mRNA independently of p53.We show that Wig-1 binds to FAS mRNA 3'-UTR and decreases its stability through an ARE in the 3'-UTR.

View Article: PubMed Central - PubMed

Affiliation: Cancer Center Karolinska (CCK), Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden.

ABSTRACT
Wig-1, also known as ZMAT3, is a p53 target gene that encodes an RNA-binding zinc-finger protein involved in the regulation of mRNA stability through binding to AU-rich elements (AREs). We have used microarray analysis to identify novel Wig-1 target mRNAs. We identified 2447 transcripts with >fourfold differential expression between Wig-1 and control small interfering (si)RNA-treated HCT116 cells. Several p53 target genes were among the deregulated transcripts. We found that Wig-1 regulates FAS and 14-3-3σ mRNA independently of p53. We show that Wig-1 binds to FAS mRNA 3'-UTR and decreases its stability through an ARE in the 3'-UTR. Depletion of Wig-1 was associated with increased cell death and reduced cell cycle arrest upon DNA damage. Our results suggest a role of Wig-1 as a survival factor that directs the p53 stress response toward cell cycle arrest rather than apoptosis through the regulation of FAS and 14-3-3σ mRNA levels.

Show MeSH
Related in: MedlinePlus