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Significant overexpression of DVL1 in Taiwanese colorectal cancer patients with liver metastasis.

Huang MY, Yen LC, Liu HC, Liu PP, Chung FY, Wang TN, Wang JY, Lin SR - Int J Mol Sci (2013)

Bottom Line: Microarray and bioinformatics analysis of 10 CRC cancer tissue specimens compared with normal adjacent tissues revealed that 31 genes were up-regulated (gene expression ratio of cancer tissue to paired normal tissue > 2) in the cancer patients.Using univariate analysis and multivariate proportional hazards regression analysis, we found that DVL1 mRNA overexpression had a significant, independent predictive value for liver metastasis in CRC patients (OR: 5.764; 95% CI: 2.588-12.837; p < 0.0001 on univariate analysis; OR: 3.768; 95% CI: 1.469-9.665; p = 0.006 on multivariate analysis).Our experimental results demonstrated that DVL1 is significantly overexpressed in CRC patients with liver metastasis, leading us to conclude that DVL1 could be a potential prognostic and predictive marker for CRC patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan. cy614112@ms14.hinet.net.

ABSTRACT
Undetected micrometastasis plays a key role in the metastasis of cancer in colorectal cancer (CRC) patients. The aim of this study is to identify a biomarker of CRC patients with liver metastasis through the detection of circulating tumor cells (CTCs). Microarray and bioinformatics analysis of 10 CRC cancer tissue specimens compared with normal adjacent tissues revealed that 31 genes were up-regulated (gene expression ratio of cancer tissue to paired normal tissue > 2) in the cancer patients. We used a weighted enzymatic chip array (WEnCA) including 31 prognosis-related genes to investigate CTCs in 214 postoperative stage I-III CRC patients and to analyze the correlation between gene expression and clinico-pathological parameters. We employed the immunohistochemistry (IHC) method with polyclonal mouse antibody against DVL1 to detect DVL1 expression in 60 CRC patients. CRC liver metastasis occurred in 19.16% (41/214) of the patients. Using univariate analysis and multivariate proportional hazards regression analysis, we found that DVL1 mRNA overexpression had a significant, independent predictive value for liver metastasis in CRC patients (OR: 5.764; 95% CI: 2.588-12.837; p < 0.0001 on univariate analysis; OR: 3.768; 95% CI: 1.469-9.665; p = 0.006 on multivariate analysis). IHC staining of the immunoreactivity of DVL1 showed that DVL1 was localized in the cytoplasm of CRC cells. High expression of DVL1 was observed in 55% (33/60) of CRC tumor specimens and was associated significantly with tumor depth, perineural invasion and liver metastasis status (all p < 0.05). Our experimental results demonstrated that DVL1 is significantly overexpressed in CRC patients with liver metastasis, leading us to conclude that DVL1 could be a potential prognostic and predictive marker for CRC patients.

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(A) The schematic representation of a weighted enzymatic chip array (WEnCA) with 31 candidate genes, one positive control (β-actin), one negative control (Oryza sativa sequence), and the blank control (dd water). Oligonucleotide fragments are blotted on membranes in triplicate. The expression levels of each gene spot were quantified then normalized based on reference gene (β-actin) density. We defined an overexpressed gene spot as occurring when the normalized spot density was 2 or more. Results of WEnCA of CRC with liver metastasis (B) and without liver metastasis (C). Circle: positive control.
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f2-ijms-14-20492: (A) The schematic representation of a weighted enzymatic chip array (WEnCA) with 31 candidate genes, one positive control (β-actin), one negative control (Oryza sativa sequence), and the blank control (dd water). Oligonucleotide fragments are blotted on membranes in triplicate. The expression levels of each gene spot were quantified then normalized based on reference gene (β-actin) density. We defined an overexpressed gene spot as occurring when the normalized spot density was 2 or more. Results of WEnCA of CRC with liver metastasis (B) and without liver metastasis (C). Circle: positive control.

Mentions: First-strand cDNA targets for hybridization were made by reverse transcription of the mRNA from the tumor and corresponding normal tissues of LARC patients in the presence of Biotin-labeled UTP by using a GeneCling® Enzymatic Gene Chip Detection Kit. The hybridized arrays were then scanned with an Epson Perfection 1670 flatbed scanner (SEIKO EPSON Corp., Nagano-ken, Japan). Subsequent quantification analysis of the intensity of each spot was carried out using AlphaEase® FC software (Alpha Innotech Corp., San Leandro, CA, USA). Spots consistently carrying a factor of two or more were considered to be differentially expressed. A deformable template extracted the gene spots and quantified their expression levels by determining the integrated intensity of each spot after background subtraction. The fold ratio for each gene was calculated as follows: spot intensity ratio = mean intensity of target gene/mean intensity of β-actin. Figure 2 provides the schematic representation of the membrane array with 31 candidate genes, one positive control (β-actin), one negative control (Oryza sativa sequence), and the blank control (dd water).


Significant overexpression of DVL1 in Taiwanese colorectal cancer patients with liver metastasis.

Huang MY, Yen LC, Liu HC, Liu PP, Chung FY, Wang TN, Wang JY, Lin SR - Int J Mol Sci (2013)

(A) The schematic representation of a weighted enzymatic chip array (WEnCA) with 31 candidate genes, one positive control (β-actin), one negative control (Oryza sativa sequence), and the blank control (dd water). Oligonucleotide fragments are blotted on membranes in triplicate. The expression levels of each gene spot were quantified then normalized based on reference gene (β-actin) density. We defined an overexpressed gene spot as occurring when the normalized spot density was 2 or more. Results of WEnCA of CRC with liver metastasis (B) and without liver metastasis (C). Circle: positive control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3821627&req=5

f2-ijms-14-20492: (A) The schematic representation of a weighted enzymatic chip array (WEnCA) with 31 candidate genes, one positive control (β-actin), one negative control (Oryza sativa sequence), and the blank control (dd water). Oligonucleotide fragments are blotted on membranes in triplicate. The expression levels of each gene spot were quantified then normalized based on reference gene (β-actin) density. We defined an overexpressed gene spot as occurring when the normalized spot density was 2 or more. Results of WEnCA of CRC with liver metastasis (B) and without liver metastasis (C). Circle: positive control.
Mentions: First-strand cDNA targets for hybridization were made by reverse transcription of the mRNA from the tumor and corresponding normal tissues of LARC patients in the presence of Biotin-labeled UTP by using a GeneCling® Enzymatic Gene Chip Detection Kit. The hybridized arrays were then scanned with an Epson Perfection 1670 flatbed scanner (SEIKO EPSON Corp., Nagano-ken, Japan). Subsequent quantification analysis of the intensity of each spot was carried out using AlphaEase® FC software (Alpha Innotech Corp., San Leandro, CA, USA). Spots consistently carrying a factor of two or more were considered to be differentially expressed. A deformable template extracted the gene spots and quantified their expression levels by determining the integrated intensity of each spot after background subtraction. The fold ratio for each gene was calculated as follows: spot intensity ratio = mean intensity of target gene/mean intensity of β-actin. Figure 2 provides the schematic representation of the membrane array with 31 candidate genes, one positive control (β-actin), one negative control (Oryza sativa sequence), and the blank control (dd water).

Bottom Line: Microarray and bioinformatics analysis of 10 CRC cancer tissue specimens compared with normal adjacent tissues revealed that 31 genes were up-regulated (gene expression ratio of cancer tissue to paired normal tissue > 2) in the cancer patients.Using univariate analysis and multivariate proportional hazards regression analysis, we found that DVL1 mRNA overexpression had a significant, independent predictive value for liver metastasis in CRC patients (OR: 5.764; 95% CI: 2.588-12.837; p < 0.0001 on univariate analysis; OR: 3.768; 95% CI: 1.469-9.665; p = 0.006 on multivariate analysis).Our experimental results demonstrated that DVL1 is significantly overexpressed in CRC patients with liver metastasis, leading us to conclude that DVL1 could be a potential prognostic and predictive marker for CRC patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Radiation Oncology, Cancer Center, Kaohsiung Medical University Hospital, Kaohsiung 807, Taiwan. cy614112@ms14.hinet.net.

ABSTRACT
Undetected micrometastasis plays a key role in the metastasis of cancer in colorectal cancer (CRC) patients. The aim of this study is to identify a biomarker of CRC patients with liver metastasis through the detection of circulating tumor cells (CTCs). Microarray and bioinformatics analysis of 10 CRC cancer tissue specimens compared with normal adjacent tissues revealed that 31 genes were up-regulated (gene expression ratio of cancer tissue to paired normal tissue > 2) in the cancer patients. We used a weighted enzymatic chip array (WEnCA) including 31 prognosis-related genes to investigate CTCs in 214 postoperative stage I-III CRC patients and to analyze the correlation between gene expression and clinico-pathological parameters. We employed the immunohistochemistry (IHC) method with polyclonal mouse antibody against DVL1 to detect DVL1 expression in 60 CRC patients. CRC liver metastasis occurred in 19.16% (41/214) of the patients. Using univariate analysis and multivariate proportional hazards regression analysis, we found that DVL1 mRNA overexpression had a significant, independent predictive value for liver metastasis in CRC patients (OR: 5.764; 95% CI: 2.588-12.837; p < 0.0001 on univariate analysis; OR: 3.768; 95% CI: 1.469-9.665; p = 0.006 on multivariate analysis). IHC staining of the immunoreactivity of DVL1 showed that DVL1 was localized in the cytoplasm of CRC cells. High expression of DVL1 was observed in 55% (33/60) of CRC tumor specimens and was associated significantly with tumor depth, perineural invasion and liver metastasis status (all p < 0.05). Our experimental results demonstrated that DVL1 is significantly overexpressed in CRC patients with liver metastasis, leading us to conclude that DVL1 could be a potential prognostic and predictive marker for CRC patients.

Show MeSH
Related in: MedlinePlus