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Up-regulation of S100A16 expression promotes epithelial-mesenchymal transition via Notch1 pathway in breast cancer.

Zhou W, Pan H, Xia T, Xue J, Cheng L, Fan P, Zhang Y, Zhu W, Xue Y, Liu X, Ding Q, Liu Y, Wang S - J. Biomed. Sci. (2014)

Bottom Line: One major mechanistic change was that S100A16 was able to up-regulate the transcription factors Notch1, ZEB1, and ZEB2, which had the capacities to directly repress the expression of epithelial markers E-cadherin and β-catenin but increase mesenchymal markers N-cadherin and vimentin, a characterized phenotype of epithelial-mensenchymal transition (EMT).In addition to display with morphologic change, migration and invasion were increased in S100A16 over-expressed MCF-7 cells.Importantly, knockdown of Notch1 by specific siRNA could reverse the EMT induced by S100A16 overexpression, which confirmed that Notch1 played a critical role in the process of EMT induced by S100A16.

View Article: PubMed Central - PubMed

Affiliation: Department of Breast Surgery, The First Affiliated Hospital with Nanjing Medical University, 300 Guangzhou Road, 210029, Nanjing, China. zhouwenbin@njmu.edu.cn.

ABSTRACT

Background: Our previous studies demonstrated that S100A16 promotes adipogenesis and is involved in weight gain attenuation induced by dietary calcium. Till now, the function of S100A16 in the breast cancer remains to be elucidated.

Results: In this study, we observed that S100A16 was expressed in higher levels in human breast cancer tissues compared with paired adjacent non-cancerous tissues. Further examination showed that overexpression of S100A16 in MCF-7 cells could increase cell proliferation and colony formation. One major mechanistic change was that S100A16 was able to up-regulate the transcription factors Notch1, ZEB1, and ZEB2, which had the capacities to directly repress the expression of epithelial markers E-cadherin and β-catenin but increase mesenchymal markers N-cadherin and vimentin, a characterized phenotype of epithelial-mensenchymal transition (EMT). In addition to display with morphologic change, migration and invasion were increased in S100A16 over-expressed MCF-7 cells. Importantly, knockdown of Notch1 by specific siRNA could reverse the EMT induced by S100A16 overexpression, which confirmed that Notch1 played a critical role in the process of EMT induced by S100A16.

Conclusions: All together, our data indicated that S100A16 had a potential function to regulate some embryonic transcription factors to promote EMT in breast cancer cells which may be an important target site for the therapy of breast cancer.

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Related in: MedlinePlus

Overexpression of S100A16 promoted EMT in MCF-7 cells. (A) Compared with control cells (MCF7-GFP), MCF7-S100A16 cells showed spindle-like, fibroblastic morphology (10×). (B, C) qRT-PCR (B) and western blot (C) analyses showed that epithelial markers E-cadherin and β-Catenin were significantly reduced in mRNA and protein levels in MCF7-S100A16 cells compared with MCF7-GFP cells, and mesenchymal markers Vimentin and N-cadherin were significantly up-regulated in MCF7-S100A16 cells (Bars, mean ± SD, P < 0.05). (D, E) Immunofluorescence staining was used to examine the location of epithelial and mesenchymal markers. After fixation, the celluar location of E-cadherin (red), β-catenin (red), Vimentin (red) and N-cadherin (red) were analyzed by confocal microscopy. Cell nuclei were stained with DAPI (4′, 6-diamidino-2-phenylindole, blue). Immunofluorescence staining showed that both E-cadherin (red) and β-catenin (red) resided in the cell membrane and intensity of fluorescence were reduced in MCF7-S100A16 cells compared with MCF7-GFP cells (D), whereas the mesenchymal markers Vimentin (red) and N-cadherin (red) were increased by S100A16 (E).
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Fig3: Overexpression of S100A16 promoted EMT in MCF-7 cells. (A) Compared with control cells (MCF7-GFP), MCF7-S100A16 cells showed spindle-like, fibroblastic morphology (10×). (B, C) qRT-PCR (B) and western blot (C) analyses showed that epithelial markers E-cadherin and β-Catenin were significantly reduced in mRNA and protein levels in MCF7-S100A16 cells compared with MCF7-GFP cells, and mesenchymal markers Vimentin and N-cadherin were significantly up-regulated in MCF7-S100A16 cells (Bars, mean ± SD, P < 0.05). (D, E) Immunofluorescence staining was used to examine the location of epithelial and mesenchymal markers. After fixation, the celluar location of E-cadherin (red), β-catenin (red), Vimentin (red) and N-cadherin (red) were analyzed by confocal microscopy. Cell nuclei were stained with DAPI (4′, 6-diamidino-2-phenylindole, blue). Immunofluorescence staining showed that both E-cadherin (red) and β-catenin (red) resided in the cell membrane and intensity of fluorescence were reduced in MCF7-S100A16 cells compared with MCF7-GFP cells (D), whereas the mesenchymal markers Vimentin (red) and N-cadherin (red) were increased by S100A16 (E).

Mentions: S100A16 over-expressed MCF-7 cells displayed morphologic changes with a spindle-like shape (Figure 3A) (similar changes were founded for T47D-S100A16 cells, data not shown). Up-regulation of S100A16 also disrupted E-cadherin mediated cell-cell adhesion. We observed that epithelial markers E-cadherin and β-Catenin were significantly reduced in mRNA and protein levels in MCF7-S100A16 cells compared with MCF7-GFP cells (Figure 3B and C) and T47D-GFP cells (Additional file 1: Figure S2). In contrast, mesenchymal markers Vimentin and N-cadherin were significantly up-regulated in MCF7-S100A16 cells (Figure 3B and C) and T47D-S100A16 cells (Additional file 1: Figure S2), indicating a characterized feature of EMT.Figure 3


Up-regulation of S100A16 expression promotes epithelial-mesenchymal transition via Notch1 pathway in breast cancer.

Zhou W, Pan H, Xia T, Xue J, Cheng L, Fan P, Zhang Y, Zhu W, Xue Y, Liu X, Ding Q, Liu Y, Wang S - J. Biomed. Sci. (2014)

Overexpression of S100A16 promoted EMT in MCF-7 cells. (A) Compared with control cells (MCF7-GFP), MCF7-S100A16 cells showed spindle-like, fibroblastic morphology (10×). (B, C) qRT-PCR (B) and western blot (C) analyses showed that epithelial markers E-cadherin and β-Catenin were significantly reduced in mRNA and protein levels in MCF7-S100A16 cells compared with MCF7-GFP cells, and mesenchymal markers Vimentin and N-cadherin were significantly up-regulated in MCF7-S100A16 cells (Bars, mean ± SD, P < 0.05). (D, E) Immunofluorescence staining was used to examine the location of epithelial and mesenchymal markers. After fixation, the celluar location of E-cadherin (red), β-catenin (red), Vimentin (red) and N-cadherin (red) were analyzed by confocal microscopy. Cell nuclei were stained with DAPI (4′, 6-diamidino-2-phenylindole, blue). Immunofluorescence staining showed that both E-cadherin (red) and β-catenin (red) resided in the cell membrane and intensity of fluorescence were reduced in MCF7-S100A16 cells compared with MCF7-GFP cells (D), whereas the mesenchymal markers Vimentin (red) and N-cadherin (red) were increased by S100A16 (E).
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Related In: Results  -  Collection

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Fig3: Overexpression of S100A16 promoted EMT in MCF-7 cells. (A) Compared with control cells (MCF7-GFP), MCF7-S100A16 cells showed spindle-like, fibroblastic morphology (10×). (B, C) qRT-PCR (B) and western blot (C) analyses showed that epithelial markers E-cadherin and β-Catenin were significantly reduced in mRNA and protein levels in MCF7-S100A16 cells compared with MCF7-GFP cells, and mesenchymal markers Vimentin and N-cadherin were significantly up-regulated in MCF7-S100A16 cells (Bars, mean ± SD, P < 0.05). (D, E) Immunofluorescence staining was used to examine the location of epithelial and mesenchymal markers. After fixation, the celluar location of E-cadherin (red), β-catenin (red), Vimentin (red) and N-cadherin (red) were analyzed by confocal microscopy. Cell nuclei were stained with DAPI (4′, 6-diamidino-2-phenylindole, blue). Immunofluorescence staining showed that both E-cadherin (red) and β-catenin (red) resided in the cell membrane and intensity of fluorescence were reduced in MCF7-S100A16 cells compared with MCF7-GFP cells (D), whereas the mesenchymal markers Vimentin (red) and N-cadherin (red) were increased by S100A16 (E).
Mentions: S100A16 over-expressed MCF-7 cells displayed morphologic changes with a spindle-like shape (Figure 3A) (similar changes were founded for T47D-S100A16 cells, data not shown). Up-regulation of S100A16 also disrupted E-cadherin mediated cell-cell adhesion. We observed that epithelial markers E-cadherin and β-Catenin were significantly reduced in mRNA and protein levels in MCF7-S100A16 cells compared with MCF7-GFP cells (Figure 3B and C) and T47D-GFP cells (Additional file 1: Figure S2). In contrast, mesenchymal markers Vimentin and N-cadherin were significantly up-regulated in MCF7-S100A16 cells (Figure 3B and C) and T47D-S100A16 cells (Additional file 1: Figure S2), indicating a characterized feature of EMT.Figure 3

Bottom Line: One major mechanistic change was that S100A16 was able to up-regulate the transcription factors Notch1, ZEB1, and ZEB2, which had the capacities to directly repress the expression of epithelial markers E-cadherin and β-catenin but increase mesenchymal markers N-cadherin and vimentin, a characterized phenotype of epithelial-mensenchymal transition (EMT).In addition to display with morphologic change, migration and invasion were increased in S100A16 over-expressed MCF-7 cells.Importantly, knockdown of Notch1 by specific siRNA could reverse the EMT induced by S100A16 overexpression, which confirmed that Notch1 played a critical role in the process of EMT induced by S100A16.

View Article: PubMed Central - PubMed

Affiliation: Department of Breast Surgery, The First Affiliated Hospital with Nanjing Medical University, 300 Guangzhou Road, 210029, Nanjing, China. zhouwenbin@njmu.edu.cn.

ABSTRACT

Background: Our previous studies demonstrated that S100A16 promotes adipogenesis and is involved in weight gain attenuation induced by dietary calcium. Till now, the function of S100A16 in the breast cancer remains to be elucidated.

Results: In this study, we observed that S100A16 was expressed in higher levels in human breast cancer tissues compared with paired adjacent non-cancerous tissues. Further examination showed that overexpression of S100A16 in MCF-7 cells could increase cell proliferation and colony formation. One major mechanistic change was that S100A16 was able to up-regulate the transcription factors Notch1, ZEB1, and ZEB2, which had the capacities to directly repress the expression of epithelial markers E-cadherin and β-catenin but increase mesenchymal markers N-cadherin and vimentin, a characterized phenotype of epithelial-mensenchymal transition (EMT). In addition to display with morphologic change, migration and invasion were increased in S100A16 over-expressed MCF-7 cells. Importantly, knockdown of Notch1 by specific siRNA could reverse the EMT induced by S100A16 overexpression, which confirmed that Notch1 played a critical role in the process of EMT induced by S100A16.

Conclusions: All together, our data indicated that S100A16 had a potential function to regulate some embryonic transcription factors to promote EMT in breast cancer cells which may be an important target site for the therapy of breast cancer.

Show MeSH
Related in: MedlinePlus