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COUP-TFII regulates metastasis of colorectal adenocarcinoma cells by modulating Snail1.

Bao Y, Gu D, Feng W, Sun X, Wang X, Zhang X, Shi Q, Cui G, Yu H, Tang C, Deng A - Br. J. Cancer (2014)

Bottom Line: Overexpression of COUP-TFII was required for cancer cells to metastasise in vivo.Chicken ovalbumin upstream promoter-transcription factor II regulated the transcription and expression of Snail1 by directly targeting the Snail1 promoter and regulated associated genes.Chicken ovalbumin upstream promoter-transcription factor II was found to be a biomarker associated with patient survival and colorectal cancer metastasis.

View Article: PubMed Central - PubMed

Affiliation: First Affiliated Hospital, Huzhou Teachers College, the First People's Hospital of Huzhou, Huzhou 313000, China.

ABSTRACT

Background: Chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII, also known as NR2F2) promotes metastasis by functioning in the tumour microenvironment; however, the role of COUP-TFII in colorectal cancer remains unknown.

Methods: Human colon adenocarcinoma tissues were collected to test COUP-TFII expression. Wound-healing and cell invasion assay were used to evaluate migration and invasion of cells. Chicken ovalbumin upstream promoter-transcription factor II and related protein expression was assessed by immunostaining, immunoblotting and real-time PCR assay. Tamoxifen-inducible COUP-TFII knockout mice were employed to test COUP-TFII functions on colon cancer metastasis in vivo.

Results: Elevated expression of COUP-TFII in colorectal adenocarcinoma tissue correlated with overexpression of the Snail1 transcription factor. High COUP-TFII expression correlated with metastasis and shorter patient survival. Chicken ovalbumin upstream promoter-transcription factor II regulated the migration and invasion of cancer cells. With Snail1, COUP-TFII inhibited expression of adherence molecules such as ZO-1, E-cadherin and β-catenin in colorectal cancer cells. Overexpression of COUP-TFII was required for cancer cells to metastasise in vivo. Chicken ovalbumin upstream promoter-transcription factor II regulated the transcription and expression of Snail1 by directly targeting the Snail1 promoter and regulated associated genes.

Conclusions: Chicken ovalbumin upstream promoter-transcription factor II was crucial for colorectal cancer metastasis and regulated cell migration and metastasis in conjunction with Snail1. Chicken ovalbumin upstream promoter-transcription factor II was found to be a biomarker associated with patient survival and colorectal cancer metastasis.

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Related in: MedlinePlus

Chicken ovalbumin upstream promoter-transcription factor II positively regulates Snail-1 expression through direct transcriptional activation. (A and B) Extracts of LOVO cells stably expressing control vector (CON), shRNA against COUP-TFII (COUP-TFII KD), or COUP-TFII-overexpressing vector (COUP-TFII OV) were examined by chromatin immunoprecipitation with anti-COUP-TFII antibody or non-immune IgG and real-time PCR of Snail-1 promoter regions containing or lacking high-affinity COUP-TFII-binding sites (black and white boxes in the map). Supershift assay was used by using antibody against COUP-TFII. −, control without antibody; antibody, with antibody against COUP-TFII (C) EMSA using LOVO cells stably expressing control vector (CON), COUP-TFII shRNA (COUP-TFII KD), or COUP-TFII-overexpressing vector (COUP-TFII OV) and a FAM-labelled oligonucleotide with the COUP-TFII-binding site from Snail-1. For supershift assay, anti-COUP-TFII antibody. (D) After ChIP with anti-Snail-1 using transfected LOVO cells as in C, target genes were detected by quantitative real-time PCR.
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fig6: Chicken ovalbumin upstream promoter-transcription factor II positively regulates Snail-1 expression through direct transcriptional activation. (A and B) Extracts of LOVO cells stably expressing control vector (CON), shRNA against COUP-TFII (COUP-TFII KD), or COUP-TFII-overexpressing vector (COUP-TFII OV) were examined by chromatin immunoprecipitation with anti-COUP-TFII antibody or non-immune IgG and real-time PCR of Snail-1 promoter regions containing or lacking high-affinity COUP-TFII-binding sites (black and white boxes in the map). Supershift assay was used by using antibody against COUP-TFII. −, control without antibody; antibody, with antibody against COUP-TFII (C) EMSA using LOVO cells stably expressing control vector (CON), COUP-TFII shRNA (COUP-TFII KD), or COUP-TFII-overexpressing vector (COUP-TFII OV) and a FAM-labelled oligonucleotide with the COUP-TFII-binding site from Snail-1. For supershift assay, anti-COUP-TFII antibody. (D) After ChIP with anti-Snail-1 using transfected LOVO cells as in C, target genes were detected by quantitative real-time PCR.

Mentions: Since COUP-TFII enhanced Snail1 expression at both the mRNA and protein levels, we tested whether Snail1 was a direct target of COUP-TFII. Chromatin immunoprecipitation assays showed binding of COUP-TFII to two distinct regions of the Snail1 promoter harbouring COUP-TFII-binding sites. Binding was abolished by knockdown of COUP-TFII, but enhanced by overexpression of COUP-TFII (Figure 6A and B).


COUP-TFII regulates metastasis of colorectal adenocarcinoma cells by modulating Snail1.

Bao Y, Gu D, Feng W, Sun X, Wang X, Zhang X, Shi Q, Cui G, Yu H, Tang C, Deng A - Br. J. Cancer (2014)

Chicken ovalbumin upstream promoter-transcription factor II positively regulates Snail-1 expression through direct transcriptional activation. (A and B) Extracts of LOVO cells stably expressing control vector (CON), shRNA against COUP-TFII (COUP-TFII KD), or COUP-TFII-overexpressing vector (COUP-TFII OV) were examined by chromatin immunoprecipitation with anti-COUP-TFII antibody or non-immune IgG and real-time PCR of Snail-1 promoter regions containing or lacking high-affinity COUP-TFII-binding sites (black and white boxes in the map). Supershift assay was used by using antibody against COUP-TFII. −, control without antibody; antibody, with antibody against COUP-TFII (C) EMSA using LOVO cells stably expressing control vector (CON), COUP-TFII shRNA (COUP-TFII KD), or COUP-TFII-overexpressing vector (COUP-TFII OV) and a FAM-labelled oligonucleotide with the COUP-TFII-binding site from Snail-1. For supershift assay, anti-COUP-TFII antibody. (D) After ChIP with anti-Snail-1 using transfected LOVO cells as in C, target genes were detected by quantitative real-time PCR.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150277&req=5

fig6: Chicken ovalbumin upstream promoter-transcription factor II positively regulates Snail-1 expression through direct transcriptional activation. (A and B) Extracts of LOVO cells stably expressing control vector (CON), shRNA against COUP-TFII (COUP-TFII KD), or COUP-TFII-overexpressing vector (COUP-TFII OV) were examined by chromatin immunoprecipitation with anti-COUP-TFII antibody or non-immune IgG and real-time PCR of Snail-1 promoter regions containing or lacking high-affinity COUP-TFII-binding sites (black and white boxes in the map). Supershift assay was used by using antibody against COUP-TFII. −, control without antibody; antibody, with antibody against COUP-TFII (C) EMSA using LOVO cells stably expressing control vector (CON), COUP-TFII shRNA (COUP-TFII KD), or COUP-TFII-overexpressing vector (COUP-TFII OV) and a FAM-labelled oligonucleotide with the COUP-TFII-binding site from Snail-1. For supershift assay, anti-COUP-TFII antibody. (D) After ChIP with anti-Snail-1 using transfected LOVO cells as in C, target genes were detected by quantitative real-time PCR.
Mentions: Since COUP-TFII enhanced Snail1 expression at both the mRNA and protein levels, we tested whether Snail1 was a direct target of COUP-TFII. Chromatin immunoprecipitation assays showed binding of COUP-TFII to two distinct regions of the Snail1 promoter harbouring COUP-TFII-binding sites. Binding was abolished by knockdown of COUP-TFII, but enhanced by overexpression of COUP-TFII (Figure 6A and B).

Bottom Line: Overexpression of COUP-TFII was required for cancer cells to metastasise in vivo.Chicken ovalbumin upstream promoter-transcription factor II regulated the transcription and expression of Snail1 by directly targeting the Snail1 promoter and regulated associated genes.Chicken ovalbumin upstream promoter-transcription factor II was found to be a biomarker associated with patient survival and colorectal cancer metastasis.

View Article: PubMed Central - PubMed

Affiliation: First Affiliated Hospital, Huzhou Teachers College, the First People's Hospital of Huzhou, Huzhou 313000, China.

ABSTRACT

Background: Chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII, also known as NR2F2) promotes metastasis by functioning in the tumour microenvironment; however, the role of COUP-TFII in colorectal cancer remains unknown.

Methods: Human colon adenocarcinoma tissues were collected to test COUP-TFII expression. Wound-healing and cell invasion assay were used to evaluate migration and invasion of cells. Chicken ovalbumin upstream promoter-transcription factor II and related protein expression was assessed by immunostaining, immunoblotting and real-time PCR assay. Tamoxifen-inducible COUP-TFII knockout mice were employed to test COUP-TFII functions on colon cancer metastasis in vivo.

Results: Elevated expression of COUP-TFII in colorectal adenocarcinoma tissue correlated with overexpression of the Snail1 transcription factor. High COUP-TFII expression correlated with metastasis and shorter patient survival. Chicken ovalbumin upstream promoter-transcription factor II regulated the migration and invasion of cancer cells. With Snail1, COUP-TFII inhibited expression of adherence molecules such as ZO-1, E-cadherin and β-catenin in colorectal cancer cells. Overexpression of COUP-TFII was required for cancer cells to metastasise in vivo. Chicken ovalbumin upstream promoter-transcription factor II regulated the transcription and expression of Snail1 by directly targeting the Snail1 promoter and regulated associated genes.

Conclusions: Chicken ovalbumin upstream promoter-transcription factor II was crucial for colorectal cancer metastasis and regulated cell migration and metastasis in conjunction with Snail1. Chicken ovalbumin upstream promoter-transcription factor II was found to be a biomarker associated with patient survival and colorectal cancer metastasis.

Show MeSH
Related in: MedlinePlus