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Epigenetic silencing of DACH1 induces the invasion and metastasis of gastric cancer by activating TGF-β signalling.

Yan W, Wu K, Herman JG, Brock MV, Zhou Y, Lu Y, Zhang Z, Yang Y, Guo M - J. Cell. Mol. Med. (2014)

Bottom Line: DACH1 is methylated in 63.3% (62/98) of primary GC and 38% (19/50) of adjacent non-tumour tissues, while no methylation was found in normal gastric mucosa.DACH1 suppresses GC proliferation, invasion and metastasis by inhibiting TGF-β signalling pathways both in vitro and in vivo.Epigenetic silencing DACH1 may induce GC cells' resistance to docetaxel.

View Article: PubMed Central - PubMed

Affiliation: Institute of Digestive Diseases, Chinese PLA General Hospital, Beijing, China.

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Related in: MedlinePlus

Effect of DACH1 on TGF-β signalling. (A) Luciferase activity was detected in BGC823 and AGS cells transfected with empty vector, DACH1 (200 ng/well), DS (40 ng/well) or ΔDS (160 ng/well), with 10 ng/ml TGF-β1 addition in each group (*P < 0.05, **P < 0.01). (B) Expression of TGF-β signalling downstream genes in DACH1 -unexpressed and -re-expressed BGC823 and AGS cells detected by Western blot; (−): absence of TGF-β1, (+): presence of TGF-β1 (T-: total; p-: phosphorylated). (C) The level of T-SMAD2 and p-SMAD2 detected by Western blot before and after knocking down DACH1 in GES-1 cells.
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fig03: Effect of DACH1 on TGF-β signalling. (A) Luciferase activity was detected in BGC823 and AGS cells transfected with empty vector, DACH1 (200 ng/well), DS (40 ng/well) or ΔDS (160 ng/well), with 10 ng/ml TGF-β1 addition in each group (*P < 0.05, **P < 0.01). (B) Expression of TGF-β signalling downstream genes in DACH1 -unexpressed and -re-expressed BGC823 and AGS cells detected by Western blot; (−): absence of TGF-β1, (+): presence of TGF-β1 (T-: total; p-: phosphorylated). (C) The level of T-SMAD2 and p-SMAD2 detected by Western blot before and after knocking down DACH1 in GES-1 cells.

Mentions: DACH1 protein contains two domains: DachBox-N and DachBox-C. Human DachBox-N shares approximately 28% amino acid identity with the SKI/SNO proteins, inhibitors of TGF-β signalling. Thus, the DachBox-N domain is also known as SKI/SNO (DS) domain [25,26]. TGF-β signalling was inhibited by DACH1 in breast and colorectal cancer by DS domain [18,19]. The function of the full-length DACH1, DS (DACH1 fragment, which encodes Smad-binding proteins' domain) and ΔDS (fragment of DACH1 deleted DS) was analysed by Dual-Luciferase reporter assay. SBE4 luciferase activity was inhibited by full-length DACH1 and DS significantly in both BGC823 and AGS cells (P < 0.05, Fig. 3A), but the activity of SBE4 has no changes before and after ΔDS transfection (P > 0.05). The results demonstrate that DS domain is critical for DACH1 function in TGF-β signalling in GC.


Epigenetic silencing of DACH1 induces the invasion and metastasis of gastric cancer by activating TGF-β signalling.

Yan W, Wu K, Herman JG, Brock MV, Zhou Y, Lu Y, Zhang Z, Yang Y, Guo M - J. Cell. Mol. Med. (2014)

Effect of DACH1 on TGF-β signalling. (A) Luciferase activity was detected in BGC823 and AGS cells transfected with empty vector, DACH1 (200 ng/well), DS (40 ng/well) or ΔDS (160 ng/well), with 10 ng/ml TGF-β1 addition in each group (*P < 0.05, **P < 0.01). (B) Expression of TGF-β signalling downstream genes in DACH1 -unexpressed and -re-expressed BGC823 and AGS cells detected by Western blot; (−): absence of TGF-β1, (+): presence of TGF-β1 (T-: total; p-: phosphorylated). (C) The level of T-SMAD2 and p-SMAD2 detected by Western blot before and after knocking down DACH1 in GES-1 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig03: Effect of DACH1 on TGF-β signalling. (A) Luciferase activity was detected in BGC823 and AGS cells transfected with empty vector, DACH1 (200 ng/well), DS (40 ng/well) or ΔDS (160 ng/well), with 10 ng/ml TGF-β1 addition in each group (*P < 0.05, **P < 0.01). (B) Expression of TGF-β signalling downstream genes in DACH1 -unexpressed and -re-expressed BGC823 and AGS cells detected by Western blot; (−): absence of TGF-β1, (+): presence of TGF-β1 (T-: total; p-: phosphorylated). (C) The level of T-SMAD2 and p-SMAD2 detected by Western blot before and after knocking down DACH1 in GES-1 cells.
Mentions: DACH1 protein contains two domains: DachBox-N and DachBox-C. Human DachBox-N shares approximately 28% amino acid identity with the SKI/SNO proteins, inhibitors of TGF-β signalling. Thus, the DachBox-N domain is also known as SKI/SNO (DS) domain [25,26]. TGF-β signalling was inhibited by DACH1 in breast and colorectal cancer by DS domain [18,19]. The function of the full-length DACH1, DS (DACH1 fragment, which encodes Smad-binding proteins' domain) and ΔDS (fragment of DACH1 deleted DS) was analysed by Dual-Luciferase reporter assay. SBE4 luciferase activity was inhibited by full-length DACH1 and DS significantly in both BGC823 and AGS cells (P < 0.05, Fig. 3A), but the activity of SBE4 has no changes before and after ΔDS transfection (P > 0.05). The results demonstrate that DS domain is critical for DACH1 function in TGF-β signalling in GC.

Bottom Line: DACH1 is methylated in 63.3% (62/98) of primary GC and 38% (19/50) of adjacent non-tumour tissues, while no methylation was found in normal gastric mucosa.DACH1 suppresses GC proliferation, invasion and metastasis by inhibiting TGF-β signalling pathways both in vitro and in vivo.Epigenetic silencing DACH1 may induce GC cells' resistance to docetaxel.

View Article: PubMed Central - PubMed

Affiliation: Institute of Digestive Diseases, Chinese PLA General Hospital, Beijing, China.

Show MeSH
Related in: MedlinePlus