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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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siRNA-mediated knockdown of NT5E affected cell migration in vitro by scratch wound healing assay. a, d GBC-SD cells transfected with pRNA-siNT5E-1. b, e GBC-SD cells transfected with unrelated sequence vector. c, f Non-transfected group. a, b, c Wound at time 0 (0 h) for GBC-SD cells incubated with control, serum-free media. d, e, f 24 hrs after wounding for SW756 cells incubated with control, serum-free media. **P < 0.01 compared to transfected with unrelated sequence vector or non-transfected group. Scale bar 100 μm
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Fig4: siRNA-mediated knockdown of NT5E affected cell migration in vitro by scratch wound healing assay. a, d GBC-SD cells transfected with pRNA-siNT5E-1. b, e GBC-SD cells transfected with unrelated sequence vector. c, f Non-transfected group. a, b, c Wound at time 0 (0 h) for GBC-SD cells incubated with control, serum-free media. d, e, f 24 hrs after wounding for SW756 cells incubated with control, serum-free media. **P < 0.01 compared to transfected with unrelated sequence vector or non-transfected group. Scale bar 100 μm

Mentions: The role of NT5E in regulating cell migration was also examined. The directed migration of GBC-SD cells transfected with pRNA- siNT5E-1, unrelated sequence vector transfected group and non-transfected group cells was compared using an in-vitro wound assay. In-vitro wound healing assays have been used with multiple cell types and are a classic and commonly used method for studying cell migration and the biology underlying it. As shown in Fig. 4, when a confluent plate of cells was scratched by a sterile pipette tip, the areas of the scratch in three groups were the same at 0 hour. After 48 hours, a small number of the GBC-SD cells transfected with pRNA -siNT5E-1 migrated to the scratch area, while the cells transfected with unrelated sequence vector pRNAi-NC and non-transfected cells migrated obviously.Fig. 4


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)

siRNA-mediated knockdown of NT5E affected cell migration in vitro by scratch wound healing assay. a, d GBC-SD cells transfected with pRNA-siNT5E-1. b, e GBC-SD cells transfected with unrelated sequence vector. c, f Non-transfected group. a, b, c Wound at time 0 (0 h) for GBC-SD cells incubated with control, serum-free media. d, e, f 24 hrs after wounding for SW756 cells incubated with control, serum-free media. **P < 0.01 compared to transfected with unrelated sequence vector or non-transfected group. Scale bar 100 μm
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig4: siRNA-mediated knockdown of NT5E affected cell migration in vitro by scratch wound healing assay. a, d GBC-SD cells transfected with pRNA-siNT5E-1. b, e GBC-SD cells transfected with unrelated sequence vector. c, f Non-transfected group. a, b, c Wound at time 0 (0 h) for GBC-SD cells incubated with control, serum-free media. d, e, f 24 hrs after wounding for SW756 cells incubated with control, serum-free media. **P < 0.01 compared to transfected with unrelated sequence vector or non-transfected group. Scale bar 100 μm
Mentions: The role of NT5E in regulating cell migration was also examined. The directed migration of GBC-SD cells transfected with pRNA- siNT5E-1, unrelated sequence vector transfected group and non-transfected group cells was compared using an in-vitro wound assay. In-vitro wound healing assays have been used with multiple cell types and are a classic and commonly used method for studying cell migration and the biology underlying it. As shown in Fig. 4, when a confluent plate of cells was scratched by a sterile pipette tip, the areas of the scratch in three groups were the same at 0 hour. After 48 hours, a small number of the GBC-SD cells transfected with pRNA -siNT5E-1 migrated to the scratch area, while the cells transfected with unrelated sequence vector pRNAi-NC and non-transfected cells migrated obviously.Fig. 4

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