Limits...
Transforming growth factor-β-induced lncRNA-Smad7 inhibits apoptosis of mouse breast cancer JygMC(A) cells.

Arase M, Horiguchi K, Ehata S, Morikawa M, Tsutsumi S, Aburatani H, Miyazono K, Koinuma D - Cancer Sci. (2014)

Bottom Line: Transforming growth factor (TGF)-β exhibits both pro-apoptotic and anti-apoptotic effects on epithelial cells in a context-dependent manner.The anti-apoptotic effect of lncRNA-Smad7 appeared to occur independently of the transcriptional regulation by TGF-β of anti-apoptotic DEC1 and pro-apoptotic Bim proteins.Small interfering RNA for lncRNA-Smad7 did not alter the process of TGF-β-induced epithelial-mesenchymal transition, phosphorylation of Smad2 or expression of the Smad7 gene, suggesting that the contribution of this lncRNA to TGF-β functions may be restricted to apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.

Show MeSH

Related in: MedlinePlus

Forced expression of lncRNA-Smad7 partially inhibits apoptosis of JygMC(A) cells. (a) Cells were infected with either adenoviral lncRNA-Smad7 expression vector (Ad-lncRNA-Smad7) or Ad-LacZ as a control. Twelve hours after infection, cells were serum starved and treated with 1 ng/mL TGF-β or 10 μM SB431542 (SB) for 48 h. Expression of lncRNA-Smad7 was determined by quantitative RT-PCR (qRT-PCR). (b) Effect of Ad-lncRNA-Smad7 on cleaved PARP level was determined by immunoblotting. Cells were treated as in (a) and lysed for SDS-PAGE. The bottom graph shows quantification of cleaved PARP normalized to α-tubulin. **P < 0.001. (c) TUNEL staining of JygMC(A) cells. Cells were treated as in (a). TUNEL, red; DAPI, blue. (d) The percentage of TUNEL-positive cells among DAPI-positive cells was determined. Error bars: standard deviations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4317863&req=5

fig04: Forced expression of lncRNA-Smad7 partially inhibits apoptosis of JygMC(A) cells. (a) Cells were infected with either adenoviral lncRNA-Smad7 expression vector (Ad-lncRNA-Smad7) or Ad-LacZ as a control. Twelve hours after infection, cells were serum starved and treated with 1 ng/mL TGF-β or 10 μM SB431542 (SB) for 48 h. Expression of lncRNA-Smad7 was determined by quantitative RT-PCR (qRT-PCR). (b) Effect of Ad-lncRNA-Smad7 on cleaved PARP level was determined by immunoblotting. Cells were treated as in (a) and lysed for SDS-PAGE. The bottom graph shows quantification of cleaved PARP normalized to α-tubulin. **P < 0.001. (c) TUNEL staining of JygMC(A) cells. Cells were treated as in (a). TUNEL, red; DAPI, blue. (d) The percentage of TUNEL-positive cells among DAPI-positive cells was determined. Error bars: standard deviations.

Mentions: The adenoviral lncRNA-Smad7 expression vector (Ad-lncRNA-Smad7) was then constructed to examine its effect on apoptosis. Expression of exogenous lncRNA-Smad7 was confirmed by qRT-PCR and compared with endogenous expression levels and with a sample without reverse transcription (Fig. 4a and data not shown). Partial inhibition by Ad-lncRNA-Smad7 of cleaved PARP expression was observed in both TGF-β-stimulated and TGF-β-unstimulated cells (Fig. 4b). The numbers of TUNEL-positive cells induced by SB431542 treatment were reduced by forced expression of lncRNA-Smad7 (Fig. 4c,d), which suggests that lncRNA-Smad7 acts as an anti-apoptotic factor downstream of TGF-β signaling.


Transforming growth factor-β-induced lncRNA-Smad7 inhibits apoptosis of mouse breast cancer JygMC(A) cells.

Arase M, Horiguchi K, Ehata S, Morikawa M, Tsutsumi S, Aburatani H, Miyazono K, Koinuma D - Cancer Sci. (2014)

Forced expression of lncRNA-Smad7 partially inhibits apoptosis of JygMC(A) cells. (a) Cells were infected with either adenoviral lncRNA-Smad7 expression vector (Ad-lncRNA-Smad7) or Ad-LacZ as a control. Twelve hours after infection, cells were serum starved and treated with 1 ng/mL TGF-β or 10 μM SB431542 (SB) for 48 h. Expression of lncRNA-Smad7 was determined by quantitative RT-PCR (qRT-PCR). (b) Effect of Ad-lncRNA-Smad7 on cleaved PARP level was determined by immunoblotting. Cells were treated as in (a) and lysed for SDS-PAGE. The bottom graph shows quantification of cleaved PARP normalized to α-tubulin. **P < 0.001. (c) TUNEL staining of JygMC(A) cells. Cells were treated as in (a). TUNEL, red; DAPI, blue. (d) The percentage of TUNEL-positive cells among DAPI-positive cells was determined. Error bars: standard deviations.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317863&req=5

fig04: Forced expression of lncRNA-Smad7 partially inhibits apoptosis of JygMC(A) cells. (a) Cells were infected with either adenoviral lncRNA-Smad7 expression vector (Ad-lncRNA-Smad7) or Ad-LacZ as a control. Twelve hours after infection, cells were serum starved and treated with 1 ng/mL TGF-β or 10 μM SB431542 (SB) for 48 h. Expression of lncRNA-Smad7 was determined by quantitative RT-PCR (qRT-PCR). (b) Effect of Ad-lncRNA-Smad7 on cleaved PARP level was determined by immunoblotting. Cells were treated as in (a) and lysed for SDS-PAGE. The bottom graph shows quantification of cleaved PARP normalized to α-tubulin. **P < 0.001. (c) TUNEL staining of JygMC(A) cells. Cells were treated as in (a). TUNEL, red; DAPI, blue. (d) The percentage of TUNEL-positive cells among DAPI-positive cells was determined. Error bars: standard deviations.
Mentions: The adenoviral lncRNA-Smad7 expression vector (Ad-lncRNA-Smad7) was then constructed to examine its effect on apoptosis. Expression of exogenous lncRNA-Smad7 was confirmed by qRT-PCR and compared with endogenous expression levels and with a sample without reverse transcription (Fig. 4a and data not shown). Partial inhibition by Ad-lncRNA-Smad7 of cleaved PARP expression was observed in both TGF-β-stimulated and TGF-β-unstimulated cells (Fig. 4b). The numbers of TUNEL-positive cells induced by SB431542 treatment were reduced by forced expression of lncRNA-Smad7 (Fig. 4c,d), which suggests that lncRNA-Smad7 acts as an anti-apoptotic factor downstream of TGF-β signaling.

Bottom Line: Transforming growth factor (TGF)-β exhibits both pro-apoptotic and anti-apoptotic effects on epithelial cells in a context-dependent manner.The anti-apoptotic effect of lncRNA-Smad7 appeared to occur independently of the transcriptional regulation by TGF-β of anti-apoptotic DEC1 and pro-apoptotic Bim proteins.Small interfering RNA for lncRNA-Smad7 did not alter the process of TGF-β-induced epithelial-mesenchymal transition, phosphorylation of Smad2 or expression of the Smad7 gene, suggesting that the contribution of this lncRNA to TGF-β functions may be restricted to apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Pathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.

Show MeSH
Related in: MedlinePlus