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Moesin-dependent cytoskeleton remodelling is associated with an anaplastic phenotype of pancreatic cancer.

Abiatari I, Esposito I, Oliveira TD, Felix K, Xin H, Penzel R, Giese T, Friess H, Kleeff J - J. Cell. Mol. Med. (2009)

Bottom Line: For functional analysis, cell growth, adhesion and invasion assays were carried out after transient and stable knock-down of moesin expression in pancreatic cancer cells.We now show that moesin knock-down increases migration, invasion and metastasis and influences extracellular matrix organization of pancreatic cancer.Moesin-regulated migratory activities of pancreatic cancer cells were in part promoted through cellular translocation of beta-catenin, and re-distribution and organization of the cytoskeleton.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, Technische Universität München, Ismaningerstrasse 22, 81675 Munich, Germany.

ABSTRACT
Cell motility is controlled by the dynamic cytoskeleton and its related proteins, such as members of the ezrin/radixin/moesin (ERM) family, which act as signalling molecules inducing cytoskeleton remodelling. Although ERM proteins have been identified as important factors in various malignancies, functional redundancy between these proteins has hindered the dissection of their individual contribution. The aim of the present study was to analyse the functional role of moesin in pancreatic malignancies. Cancer cells of different malignant lesions of human and transgenic mice pancreata were evaluated by immunohistochemistry. For functional analysis, cell growth, adhesion and invasion assays were carried out after transient and stable knock-down of moesin expression in pancreatic cancer cells. In vivo tumourigenicity was determined using orthotopic and metastatic mouse tumour models. We now show that moesin knock-down increases migration, invasion and metastasis and influences extracellular matrix organization of pancreatic cancer. Moesin-regulated migratory activities of pancreatic cancer cells were in part promoted through cellular translocation of beta-catenin, and re-distribution and organization of the cytoskeleton. Analysis of human and different transgenic mouse pancreatic cancers demonstrated that moesin is a phenotypic marker for anaplastic carcinoma, suggesting that this ERM protein plays a specific role in pancreatic carcinogenesis.

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High expression of moesin in PAC and loss of expression in PDAC. (A) Immunohistochemical staining of normal pancreas (1), PDAC (2) and PAC (3) samples using a specific ezrin antibody and of PDAC (4) and PAC (5) samples, using a specific radixin antibody (20× magnification). (B) Immunohistochemical staining of normal pancreas (1), CP (2), PDAC (3–5) and PAC (6, 7) samples using a specific moesin monoclonal antibody (20× magnification). Immunofluorescence confocal microscopy staining of PDAC (8) and PAC (9) tissues using the moesin antibody (red). Note actin (green), and nuclear (blue) DAPI counterstaining. (C) Immunohistochemical staining of PAC (1) containing PanIN lesions (2) with a specific moesin antibody (20× magnification). (3) QRT-PCR data of ezrin/radixin/moesin expression in pancreatic stellate cells (PSC) and peripheral blood mononuclear cells (PBMC). (D) QRT-PCR showing expression of ezrin/radixin/moesin (left panel), and immunoblot analysis showing expression of ezrin/radixin/moesin in eight cultured pancreatic cancer cell lines (right panel), as well as in two primary pancreatic cancer cell lines (pPDAC1; pPDAC2; lower panel).
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fig01: High expression of moesin in PAC and loss of expression in PDAC. (A) Immunohistochemical staining of normal pancreas (1), PDAC (2) and PAC (3) samples using a specific ezrin antibody and of PDAC (4) and PAC (5) samples, using a specific radixin antibody (20× magnification). (B) Immunohistochemical staining of normal pancreas (1), CP (2), PDAC (3–5) and PAC (6, 7) samples using a specific moesin monoclonal antibody (20× magnification). Immunofluorescence confocal microscopy staining of PDAC (8) and PAC (9) tissues using the moesin antibody (red). Note actin (green), and nuclear (blue) DAPI counterstaining. (C) Immunohistochemical staining of PAC (1) containing PanIN lesions (2) with a specific moesin antibody (20× magnification). (3) QRT-PCR data of ezrin/radixin/moesin expression in pancreatic stellate cells (PSC) and peripheral blood mononuclear cells (PBMC). (D) QRT-PCR showing expression of ezrin/radixin/moesin (left panel), and immunoblot analysis showing expression of ezrin/radixin/moesin in eight cultured pancreatic cancer cell lines (right panel), as well as in two primary pancreatic cancer cell lines (pPDAC1; pPDAC2; lower panel).

Mentions: Characterization of ERM proteins in pancreatic tissues revealed strong expression of Ezrin in small ducts and centroacinar cells of the normal pancreas (Fig. 1A-1) and in cancer cells of PDAC (Fig. 1A-2). Ezrin was also present in some cells of PAC (Fig. 1A-3). Expression of radixin was observed in the cancer cells of PDAC and PAC tissues, but to a comparably lesser extent (Fig. 1A-4, 5). Expression of moesin was faintly detectable in normal pancreatic tissues (Fig. 1B-1) but present in fibroblasts/stellate cells and inflammatory cells of chronic pancreatitis (CP) tissues (Fig. 1B-2). Expression of moesin (and to a lesser extent of ezrin and radixin) in cultured pancreatic stellate cells as well as in peripheral blood mononuclear cells was also confirmed by QRT-PCR (Fig. 1C-3). None of the tested PDAC tissues (n= 70) showed positive staining for moesin in the cancer cells (Fig. 1B-3–5, 8). In contrast, moesin was found to be abundant in the neoplastic cells of PAC (n= 8) (Fig. 1B-6, 7), which was also confirmed by confocal microscopy, demonstrating to some extent co-localization with actin (Fig. 1B-9).


Moesin-dependent cytoskeleton remodelling is associated with an anaplastic phenotype of pancreatic cancer.

Abiatari I, Esposito I, Oliveira TD, Felix K, Xin H, Penzel R, Giese T, Friess H, Kleeff J - J. Cell. Mol. Med. (2009)

High expression of moesin in PAC and loss of expression in PDAC. (A) Immunohistochemical staining of normal pancreas (1), PDAC (2) and PAC (3) samples using a specific ezrin antibody and of PDAC (4) and PAC (5) samples, using a specific radixin antibody (20× magnification). (B) Immunohistochemical staining of normal pancreas (1), CP (2), PDAC (3–5) and PAC (6, 7) samples using a specific moesin monoclonal antibody (20× magnification). Immunofluorescence confocal microscopy staining of PDAC (8) and PAC (9) tissues using the moesin antibody (red). Note actin (green), and nuclear (blue) DAPI counterstaining. (C) Immunohistochemical staining of PAC (1) containing PanIN lesions (2) with a specific moesin antibody (20× magnification). (3) QRT-PCR data of ezrin/radixin/moesin expression in pancreatic stellate cells (PSC) and peripheral blood mononuclear cells (PBMC). (D) QRT-PCR showing expression of ezrin/radixin/moesin (left panel), and immunoblot analysis showing expression of ezrin/radixin/moesin in eight cultured pancreatic cancer cell lines (right panel), as well as in two primary pancreatic cancer cell lines (pPDAC1; pPDAC2; lower panel).
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fig01: High expression of moesin in PAC and loss of expression in PDAC. (A) Immunohistochemical staining of normal pancreas (1), PDAC (2) and PAC (3) samples using a specific ezrin antibody and of PDAC (4) and PAC (5) samples, using a specific radixin antibody (20× magnification). (B) Immunohistochemical staining of normal pancreas (1), CP (2), PDAC (3–5) and PAC (6, 7) samples using a specific moesin monoclonal antibody (20× magnification). Immunofluorescence confocal microscopy staining of PDAC (8) and PAC (9) tissues using the moesin antibody (red). Note actin (green), and nuclear (blue) DAPI counterstaining. (C) Immunohistochemical staining of PAC (1) containing PanIN lesions (2) with a specific moesin antibody (20× magnification). (3) QRT-PCR data of ezrin/radixin/moesin expression in pancreatic stellate cells (PSC) and peripheral blood mononuclear cells (PBMC). (D) QRT-PCR showing expression of ezrin/radixin/moesin (left panel), and immunoblot analysis showing expression of ezrin/radixin/moesin in eight cultured pancreatic cancer cell lines (right panel), as well as in two primary pancreatic cancer cell lines (pPDAC1; pPDAC2; lower panel).
Mentions: Characterization of ERM proteins in pancreatic tissues revealed strong expression of Ezrin in small ducts and centroacinar cells of the normal pancreas (Fig. 1A-1) and in cancer cells of PDAC (Fig. 1A-2). Ezrin was also present in some cells of PAC (Fig. 1A-3). Expression of radixin was observed in the cancer cells of PDAC and PAC tissues, but to a comparably lesser extent (Fig. 1A-4, 5). Expression of moesin was faintly detectable in normal pancreatic tissues (Fig. 1B-1) but present in fibroblasts/stellate cells and inflammatory cells of chronic pancreatitis (CP) tissues (Fig. 1B-2). Expression of moesin (and to a lesser extent of ezrin and radixin) in cultured pancreatic stellate cells as well as in peripheral blood mononuclear cells was also confirmed by QRT-PCR (Fig. 1C-3). None of the tested PDAC tissues (n= 70) showed positive staining for moesin in the cancer cells (Fig. 1B-3–5, 8). In contrast, moesin was found to be abundant in the neoplastic cells of PAC (n= 8) (Fig. 1B-6, 7), which was also confirmed by confocal microscopy, demonstrating to some extent co-localization with actin (Fig. 1B-9).

Bottom Line: For functional analysis, cell growth, adhesion and invasion assays were carried out after transient and stable knock-down of moesin expression in pancreatic cancer cells.We now show that moesin knock-down increases migration, invasion and metastasis and influences extracellular matrix organization of pancreatic cancer.Moesin-regulated migratory activities of pancreatic cancer cells were in part promoted through cellular translocation of beta-catenin, and re-distribution and organization of the cytoskeleton.

View Article: PubMed Central - PubMed

Affiliation: Department of General Surgery, Technische Universität München, Ismaningerstrasse 22, 81675 Munich, Germany.

ABSTRACT
Cell motility is controlled by the dynamic cytoskeleton and its related proteins, such as members of the ezrin/radixin/moesin (ERM) family, which act as signalling molecules inducing cytoskeleton remodelling. Although ERM proteins have been identified as important factors in various malignancies, functional redundancy between these proteins has hindered the dissection of their individual contribution. The aim of the present study was to analyse the functional role of moesin in pancreatic malignancies. Cancer cells of different malignant lesions of human and transgenic mice pancreata were evaluated by immunohistochemistry. For functional analysis, cell growth, adhesion and invasion assays were carried out after transient and stable knock-down of moesin expression in pancreatic cancer cells. In vivo tumourigenicity was determined using orthotopic and metastatic mouse tumour models. We now show that moesin knock-down increases migration, invasion and metastasis and influences extracellular matrix organization of pancreatic cancer. Moesin-regulated migratory activities of pancreatic cancer cells were in part promoted through cellular translocation of beta-catenin, and re-distribution and organization of the cytoskeleton. Analysis of human and different transgenic mouse pancreatic cancers demonstrated that moesin is a phenotypic marker for anaplastic carcinoma, suggesting that this ERM protein plays a specific role in pancreatic carcinogenesis.

Show MeSH
Related in: MedlinePlus