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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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Generation of a cell line stably expressing moesin shRNA. (A) Immunoblot analysis showing SU8686 cells stably transduced with shRNA for moesin (Lv-MSN) as compared to moesin siRNA-transfected (MSN RNAi) and scrambled shRNA (Lv-control) cells (left) and densitometric quantification of moesin down-regulation in SU8686 cancer cells (right). Data are shown as mean ± S.E.M. (B) Immunofluorescence analysis of Lv-MSN and Lv-control transduced SU8686 cells showing moesin (red), actin (green) and nuclear (blue) counterstaining. (C) Quantification of anoikis assay for Lv-MSN and Lv-control cells after detection of cell viability by FACS. Apoptotic cells were separated from the dead cell fraction. Data are shown as mean ± S.E.M. *P < 0.05 compared to control cells. (D) Immunofluorescence analysis of actin cytoskeleton (green) using phalloidin staining, upon serum activation in Lv-control (1, 2) and Lv-MSN (4, 5) SU8686 cells. Vinculin staining (red) of Lv-control (3) and Lv-MSN (6) cells, demonstrating focal adhesion co-localization with radiating F-actin (green) stress fibre ends.
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fig04: Generation of a cell line stably expressing moesin shRNA. (A) Immunoblot analysis showing SU8686 cells stably transduced with shRNA for moesin (Lv-MSN) as compared to moesin siRNA-transfected (MSN RNAi) and scrambled shRNA (Lv-control) cells (left) and densitometric quantification of moesin down-regulation in SU8686 cancer cells (right). Data are shown as mean ± S.E.M. (B) Immunofluorescence analysis of Lv-MSN and Lv-control transduced SU8686 cells showing moesin (red), actin (green) and nuclear (blue) counterstaining. (C) Quantification of anoikis assay for Lv-MSN and Lv-control cells after detection of cell viability by FACS. Apoptotic cells were separated from the dead cell fraction. Data are shown as mean ± S.E.M. *P < 0.05 compared to control cells. (D) Immunofluorescence analysis of actin cytoskeleton (green) using phalloidin staining, upon serum activation in Lv-control (1, 2) and Lv-MSN (4, 5) SU8686 cells. Vinculin staining (red) of Lv-control (3) and Lv-MSN (6) cells, demonstrating focal adhesion co-localization with radiating F-actin (green) stress fibre ends.

Mentions: For functional assessment of moesin, transient RNAi transfection of Panc-1 and SU8686 pancreatic cancer cell lines was utilized. Immunoblot analysis demonstrated reduced moesin expression after 96 hrs of MSN RNAi compared to control RNAi (Figs 2A, 3A and 4A).


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)

Generation of a cell line stably expressing moesin shRNA. (A) Immunoblot analysis showing SU8686 cells stably transduced with shRNA for moesin (Lv-MSN) as compared to moesin siRNA-transfected (MSN RNAi) and scrambled shRNA (Lv-control) cells (left) and densitometric quantification of moesin down-regulation in SU8686 cancer cells (right). Data are shown as mean ± S.E.M. (B) Immunofluorescence analysis of Lv-MSN and Lv-control transduced SU8686 cells showing moesin (red), actin (green) and nuclear (blue) counterstaining. (C) Quantification of anoikis assay for Lv-MSN and Lv-control cells after detection of cell viability by FACS. Apoptotic cells were separated from the dead cell fraction. Data are shown as mean ± S.E.M. *P < 0.05 compared to control cells. (D) Immunofluorescence analysis of actin cytoskeleton (green) using phalloidin staining, upon serum activation in Lv-control (1, 2) and Lv-MSN (4, 5) SU8686 cells. Vinculin staining (red) of Lv-control (3) and Lv-MSN (6) cells, demonstrating focal adhesion co-localization with radiating F-actin (green) stress fibre ends.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3822753&req=5

fig04: Generation of a cell line stably expressing moesin shRNA. (A) Immunoblot analysis showing SU8686 cells stably transduced with shRNA for moesin (Lv-MSN) as compared to moesin siRNA-transfected (MSN RNAi) and scrambled shRNA (Lv-control) cells (left) and densitometric quantification of moesin down-regulation in SU8686 cancer cells (right). Data are shown as mean ± S.E.M. (B) Immunofluorescence analysis of Lv-MSN and Lv-control transduced SU8686 cells showing moesin (red), actin (green) and nuclear (blue) counterstaining. (C) Quantification of anoikis assay for Lv-MSN and Lv-control cells after detection of cell viability by FACS. Apoptotic cells were separated from the dead cell fraction. Data are shown as mean ± S.E.M. *P < 0.05 compared to control cells. (D) Immunofluorescence analysis of actin cytoskeleton (green) using phalloidin staining, upon serum activation in Lv-control (1, 2) and Lv-MSN (4, 5) SU8686 cells. Vinculin staining (red) of Lv-control (3) and Lv-MSN (6) cells, demonstrating focal adhesion co-localization with radiating F-actin (green) stress fibre ends.
Mentions: For functional assessment of moesin, transient RNAi transfection of Panc-1 and SU8686 pancreatic cancer cell lines was utilized. Immunoblot analysis demonstrated reduced moesin expression after 96 hrs of MSN RNAi compared to control RNAi (Figs 2A, 3A and 4A).

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