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DSC3 expression is regulated by p53, and methylation of DSC3 DNA is a prognostic marker in human colorectal cancer.

Cui T, Chen Y, Yang L, Knösel T, Zöller K, Huber O, Petersen I - Br. J. Cancer (2011)

Bottom Line: Methylation of DSC3 genomic sequences was found in 41% (41 out of 99) of primary CRC, being associated with poor prognosis (P=0.002).Methylation status of DSC3 DNA is a prognostic marker for CRC.P53 appears to have an important role in regulating DSC3 expression.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, University Hospital Jena, Ziegelmühlenweg 1, Jena 07743, Germany.

ABSTRACT

Background: Desmocollin 3 (DSC3), a member of the cadherin superfamily and integral component of desmosomes, is involved in carcinogenesis. However, the role of DSC3 in colorectal cancer (CRC) has not yet been established.

Methods: Desmocollin 3 expression in CRC cell lines was analysed by RT-PCR and western blotting. Methylation status of DSC3 was examined by demethylation tests, methylation-specific PCR, and bisulphite sequencing (BS). The regulatory role of p53 was investigated by transfection.

Results: Desmocollin 3 was downregulated in CRC cells at mRNA and protein levels. Desmocollin 3 expression was restored in five out of seven cell lines after 5-aza-2'-deoxycytidine (DAC) treatment. A heterogeneous methylation pattern was detected by BS in promoter region and exon 1 of DSC3. Methylation of DSC3 genomic sequences was found in 41% (41 out of 99) of primary CRC, being associated with poor prognosis (P=0.002). Transfection of p53 alone or in combination of DAC increased the DSC3 expression. Similarly, treatment with p53 inducer adriamycin alone or in combination with DAC enhanced DSC3 expression.

Conclusions: DNA methylation contributes to downregulation of DSC3 in CRC cell lines. Methylation status of DSC3 DNA is a prognostic marker for CRC. P53 appears to have an important role in regulating DSC3 expression.

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Related in: MedlinePlus

Demethylation tests in CRC cell lines. (A) Semiquantitative RT–PCR and (B) real-time RT–PCR showed that after treatment with 10 μ DAC for 96 h, DSC3 mRNA expression was upregulated. (−)=untreated; (+)=treated with DAC.
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fig2: Demethylation tests in CRC cell lines. (A) Semiquantitative RT–PCR and (B) real-time RT–PCR showed that after treatment with 10 μ DAC for 96 h, DSC3 mRNA expression was upregulated. (−)=untreated; (+)=treated with DAC.

Mentions: To explore the epigenetic regulation of DSC3, seven colon cancer cell lines including HT-29, LoVo, WiDr, SW480, CX-2, HCT116, and HRT-18 were selected for the demethylation tests. After treatment with 10 μ DAC for 96 h, DSC3 mRNA expression was restored in five (HT-29, LoVo, WiDr, HCT116, and HRT-18) out of seven cell lines. While in the other two cell lines (SW480 and CX-2), no restoration of DSC3 expression was detectable (Figures 2A and B).


DSC3 expression is regulated by p53, and methylation of DSC3 DNA is a prognostic marker in human colorectal cancer.

Cui T, Chen Y, Yang L, Knösel T, Zöller K, Huber O, Petersen I - Br. J. Cancer (2011)

Demethylation tests in CRC cell lines. (A) Semiquantitative RT–PCR and (B) real-time RT–PCR showed that after treatment with 10 μ DAC for 96 h, DSC3 mRNA expression was upregulated. (−)=untreated; (+)=treated with DAC.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3065270&req=5

fig2: Demethylation tests in CRC cell lines. (A) Semiquantitative RT–PCR and (B) real-time RT–PCR showed that after treatment with 10 μ DAC for 96 h, DSC3 mRNA expression was upregulated. (−)=untreated; (+)=treated with DAC.
Mentions: To explore the epigenetic regulation of DSC3, seven colon cancer cell lines including HT-29, LoVo, WiDr, SW480, CX-2, HCT116, and HRT-18 were selected for the demethylation tests. After treatment with 10 μ DAC for 96 h, DSC3 mRNA expression was restored in five (HT-29, LoVo, WiDr, HCT116, and HRT-18) out of seven cell lines. While in the other two cell lines (SW480 and CX-2), no restoration of DSC3 expression was detectable (Figures 2A and B).

Bottom Line: Methylation of DSC3 genomic sequences was found in 41% (41 out of 99) of primary CRC, being associated with poor prognosis (P=0.002).Methylation status of DSC3 DNA is a prognostic marker for CRC.P53 appears to have an important role in regulating DSC3 expression.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, University Hospital Jena, Ziegelmühlenweg 1, Jena 07743, Germany.

ABSTRACT

Background: Desmocollin 3 (DSC3), a member of the cadherin superfamily and integral component of desmosomes, is involved in carcinogenesis. However, the role of DSC3 in colorectal cancer (CRC) has not yet been established.

Methods: Desmocollin 3 expression in CRC cell lines was analysed by RT-PCR and western blotting. Methylation status of DSC3 was examined by demethylation tests, methylation-specific PCR, and bisulphite sequencing (BS). The regulatory role of p53 was investigated by transfection.

Results: Desmocollin 3 was downregulated in CRC cells at mRNA and protein levels. Desmocollin 3 expression was restored in five out of seven cell lines after 5-aza-2'-deoxycytidine (DAC) treatment. A heterogeneous methylation pattern was detected by BS in promoter region and exon 1 of DSC3. Methylation of DSC3 genomic sequences was found in 41% (41 out of 99) of primary CRC, being associated with poor prognosis (P=0.002). Transfection of p53 alone or in combination of DAC increased the DSC3 expression. Similarly, treatment with p53 inducer adriamycin alone or in combination with DAC enhanced DSC3 expression.

Conclusions: DNA methylation contributes to downregulation of DSC3 in CRC cell lines. Methylation status of DSC3 DNA is a prognostic marker for CRC. P53 appears to have an important role in regulating DSC3 expression.

Show MeSH
Related in: MedlinePlus