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NT5E and FcGBP as key regulators of TGF-1-induced epithelial-mesenchymal transition (EMT) are associated with tumor progression and survival of patients with gallbladder cancer.

Xiong L, Wen Y, Miao X, Yang Z - Cell Tissue Res. (2013)

Bottom Line: In the present study, we used DNA micro-array analysis to search for differentially expressed genes in the TGF-β1 induced gallbladder carcinoma cell line (GBC-SD cells), as compared with normal GBC-SD cells.NT5E (CD73) is the most increased gene, while the Fc fragment of the IgG binding protein (FcGBP) is the most decreased gene.Immunochemistry and clinicopathological results showed that the expression of NT5E and FcGBP in gallbladder adenocarcinoma is an independent marker for evaluation of the disease progression, clinical biological behaviors and prognosis.

View Article: PubMed Central - PubMed

Affiliation: Research Laboratory of Hepatobiliary Diseases, Second Xiangya Hospital, Central South University, 139# Middle Renmin road, Changsha, Hunan, 410011, China.

ABSTRACT
Epithelial-mesenchymal transitions (EMTs) are essential manifestations of epithelial cell plasticity during tumor progression. Transforming growth factor-β(TGF-β) modulates epithelial plasticity in tumor physiological contexts by inducing EMT, which is associated with the altered expression of genes. In the present study, we used DNA micro-array analysis to search for differentially expressed genes in the TGF-β1 induced gallbladder carcinoma cell line (GBC-SD cells), as compared with normal GBC-SD cells. We identified 225 differentially expressed genes, including 144 that were over-expressed and 81 that were under-expressed in the TGF-β1 induced GBC-SD cells. NT5E (CD73) is the most increased gene, while the Fc fragment of the IgG binding protein (FcGBP) is the most decreased gene. The expression patterns of these two genes in gallbladder adenocarcinoma and chronic cholecystitis tissue were consistent with the micro-array data. Immunochemistry and clinicopathological results showed that the expression of NT5E and FcGBP in gallbladder adenocarcinoma is an independent marker for evaluation of the disease progression, clinical biological behaviors and prognosis. The data from the current study indicate that differential NT5E and FcGBP expressions could be further evaluated as biomarkers for predicting survival of patients with gallbladder cancer and that NT5E and FcGBP could be promising targets in the control of gallbladder cancer progression.

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RT-PCR and Western blot analysis of the expression level of NT5E and FcGBP in the gallbladder adenocarcinoma and chronic cholecystitis samples. a, b The total RNA was both extracted from specimens of 15 patients diagnosed as having gallbladder adenocarcinoma and 15 patients diagnosed as having chronic cholecystitis. The mRNA transcription expression levels of NT5E and FcGBP were evaluated by RT-PCR. The values were normalized to GAPDH as an internal control. c, d Western blotting analysis of the levels of NT5E and FcGBP in the gallbladder adenocarcinoma and chronic cholecystitis samples. Proteins were separated by 11.5 % SDS-PAGE. The gels were electroeluted to PVDF membrane and each blot was analyzed with a different antibody: anti- NT5E and anti-FcGBP. The values were normalized to actin as an internal control. Mean expression values ± SE are shown. Asterisks indicate that the expression differences between TGF-β1 induced GBC-SD cells and normal GBC-SD cells are statistically significant (p < 0.05 for Student’s t-test)
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Fig1: RT-PCR and Western blot analysis of the expression level of NT5E and FcGBP in the gallbladder adenocarcinoma and chronic cholecystitis samples. a, b The total RNA was both extracted from specimens of 15 patients diagnosed as having gallbladder adenocarcinoma and 15 patients diagnosed as having chronic cholecystitis. The mRNA transcription expression levels of NT5E and FcGBP were evaluated by RT-PCR. The values were normalized to GAPDH as an internal control. c, d Western blotting analysis of the levels of NT5E and FcGBP in the gallbladder adenocarcinoma and chronic cholecystitis samples. Proteins were separated by 11.5 % SDS-PAGE. The gels were electroeluted to PVDF membrane and each blot was analyzed with a different antibody: anti- NT5E and anti-FcGBP. The values were normalized to actin as an internal control. Mean expression values ± SE are shown. Asterisks indicate that the expression differences between TGF-β1 induced GBC-SD cells and normal GBC-SD cells are statistically significant (p < 0.05 for Student’s t-test)

Mentions: To explore the mechanism of invasion and metastasis of gallbladder cancer, DNA microarray technology was applied to investigate the differentially expressed genes between GBC cells with TGF-β1 induced epithelial mesenchymal transition (EMT) and normal GBC cells. Results revealed by DNA microarray showed that Hex, interior label and external label were normal in positive control and negative in negative control, with an authentic repetitiveness of housekeeping genes (ratio CV ≤ 0.3) and leak rate ≤3‰, indicating that the data are valid without genomic DNA contamination. The distribution of differential gene expressions in TGF-β1 induced and control GBC-SD cells is shown in a scatter plot form in Fig. 1. These items suggested that the samples were not pollution. Two hundred and twenty-five differentially expressed genes were identified between GBC-SD cells with and without TGF-β1 induced EMT, among which 144 genes were up-regulated (Supplement Table 1) and 81 genes were down-regulated (Supplement Table 2) in GBC cells after TGF-β1 induced EMT. NT5E is the most increased gene while FcGBP is the most decreased gene.Fig. 1


NT5E and FcGBP as key regulators of TGF-1-induced epithelial-mesenchymal transition (EMT) are associated with tumor progression and survival of patients with gallbladder cancer.

Xiong L, Wen Y, Miao X, Yang Z - Cell Tissue Res. (2013)

RT-PCR and Western blot analysis of the expression level of NT5E and FcGBP in the gallbladder adenocarcinoma and chronic cholecystitis samples. a, b The total RNA was both extracted from specimens of 15 patients diagnosed as having gallbladder adenocarcinoma and 15 patients diagnosed as having chronic cholecystitis. The mRNA transcription expression levels of NT5E and FcGBP were evaluated by RT-PCR. The values were normalized to GAPDH as an internal control. c, d Western blotting analysis of the levels of NT5E and FcGBP in the gallbladder adenocarcinoma and chronic cholecystitis samples. Proteins were separated by 11.5 % SDS-PAGE. The gels were electroeluted to PVDF membrane and each blot was analyzed with a different antibody: anti- NT5E and anti-FcGBP. The values were normalized to actin as an internal control. Mean expression values ± SE are shown. Asterisks indicate that the expression differences between TGF-β1 induced GBC-SD cells and normal GBC-SD cells are statistically significant (p < 0.05 for Student’s t-test)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3921456&req=5

Fig1: RT-PCR and Western blot analysis of the expression level of NT5E and FcGBP in the gallbladder adenocarcinoma and chronic cholecystitis samples. a, b The total RNA was both extracted from specimens of 15 patients diagnosed as having gallbladder adenocarcinoma and 15 patients diagnosed as having chronic cholecystitis. The mRNA transcription expression levels of NT5E and FcGBP were evaluated by RT-PCR. The values were normalized to GAPDH as an internal control. c, d Western blotting analysis of the levels of NT5E and FcGBP in the gallbladder adenocarcinoma and chronic cholecystitis samples. Proteins were separated by 11.5 % SDS-PAGE. The gels were electroeluted to PVDF membrane and each blot was analyzed with a different antibody: anti- NT5E and anti-FcGBP. The values were normalized to actin as an internal control. Mean expression values ± SE are shown. Asterisks indicate that the expression differences between TGF-β1 induced GBC-SD cells and normal GBC-SD cells are statistically significant (p < 0.05 for Student’s t-test)
Mentions: To explore the mechanism of invasion and metastasis of gallbladder cancer, DNA microarray technology was applied to investigate the differentially expressed genes between GBC cells with TGF-β1 induced epithelial mesenchymal transition (EMT) and normal GBC cells. Results revealed by DNA microarray showed that Hex, interior label and external label were normal in positive control and negative in negative control, with an authentic repetitiveness of housekeeping genes (ratio CV ≤ 0.3) and leak rate ≤3‰, indicating that the data are valid without genomic DNA contamination. The distribution of differential gene expressions in TGF-β1 induced and control GBC-SD cells is shown in a scatter plot form in Fig. 1. These items suggested that the samples were not pollution. Two hundred and twenty-five differentially expressed genes were identified between GBC-SD cells with and without TGF-β1 induced EMT, among which 144 genes were up-regulated (Supplement Table 1) and 81 genes were down-regulated (Supplement Table 2) in GBC cells after TGF-β1 induced EMT. NT5E is the most increased gene while FcGBP is the most decreased gene.Fig. 1

Bottom Line: In the present study, we used DNA micro-array analysis to search for differentially expressed genes in the TGF-β1 induced gallbladder carcinoma cell line (GBC-SD cells), as compared with normal GBC-SD cells.NT5E (CD73) is the most increased gene, while the Fc fragment of the IgG binding protein (FcGBP) is the most decreased gene.Immunochemistry and clinicopathological results showed that the expression of NT5E and FcGBP in gallbladder adenocarcinoma is an independent marker for evaluation of the disease progression, clinical biological behaviors and prognosis.

View Article: PubMed Central - PubMed

Affiliation: Research Laboratory of Hepatobiliary Diseases, Second Xiangya Hospital, Central South University, 139# Middle Renmin road, Changsha, Hunan, 410011, China.

ABSTRACT
Epithelial-mesenchymal transitions (EMTs) are essential manifestations of epithelial cell plasticity during tumor progression. Transforming growth factor-β(TGF-β) modulates epithelial plasticity in tumor physiological contexts by inducing EMT, which is associated with the altered expression of genes. In the present study, we used DNA micro-array analysis to search for differentially expressed genes in the TGF-β1 induced gallbladder carcinoma cell line (GBC-SD cells), as compared with normal GBC-SD cells. We identified 225 differentially expressed genes, including 144 that were over-expressed and 81 that were under-expressed in the TGF-β1 induced GBC-SD cells. NT5E (CD73) is the most increased gene, while the Fc fragment of the IgG binding protein (FcGBP) is the most decreased gene. The expression patterns of these two genes in gallbladder adenocarcinoma and chronic cholecystitis tissue were consistent with the micro-array data. Immunochemistry and clinicopathological results showed that the expression of NT5E and FcGBP in gallbladder adenocarcinoma is an independent marker for evaluation of the disease progression, clinical biological behaviors and prognosis. The data from the current study indicate that differential NT5E and FcGBP expressions could be further evaluated as biomarkers for predicting survival of patients with gallbladder cancer and that NT5E and FcGBP could be promising targets in the control of gallbladder cancer progression.

Show MeSH
Related in: MedlinePlus