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Josephine C. Adams - One of the best experts on this subject based on the ideXlab platform.

  • The roles of Fascins in health and disease
    The Journal of pathology, 2011
    Co-Authors: Yosuke Hashimoto, Dae Joong Kim, Josephine C. Adams
    Abstract:

    Fascins are actin-binding proteins that cross-link filamentous actin into tightly packed parallel bundles. These bundles are important for the organization and morphology of an extremely diverse set of sub-cellular structures that include dynamic and stable cell-surface protrusions, stress fibres, and the specialized actin bundles of photoreceptor and stereocilia cells. In this review, we discuss the Fascin gene family and its evolution, the actin-bundling activity of Fascins and the molecular pathways by which it is regulated, and the role of the diverse actin/Fascin structures in normal cellular processes. We discuss the mechanisms by which Fascins contribute to disease pathologies, especially cancer, where Fascin-1 is emerging as a novel therapeutic target in carcinoma metastasis.

  • Rac regulates the interaction of Fascin with protein kinase C in cell migration.
    Journal of Cell Science, 2008
    Co-Authors: Maddy Parsons, Josephine C. Adams
    Abstract:

    Fascin is an actin-bundling protein that is low or absent in normal epithelia; its upregulation correlates with poor prognosis in many human carcinomas. We have recently demonstrated in mouse xenograft models that Fascin contributes to tumour development and metastasis through its dual actin-bundling and active PKC-binding activities. Rac was implicated as a regulator of Fascin-dependent colon carcinoma cell migration in vitro. Here, we tested the hypothesis that Rac regulates the interaction of Fascin with active PKC. The major conventional PKC in colon carcinoma cells is protein kinase Cgamma (PKCgamma). Endogenous PKCgamma, Fascin and Rac1 colocalised at lamellipodial margins of migrating cells. Colocalisation of Fascin and PKCgamma depended on Rac activity, and inhibition of Rac decreased PKCgamma activity in cell extracts but not in vitro. Fluorescence resonance energy transfer/fluorescence lifetime imaging microscopy uncovered that Fascin and PKCgamma interact in protrusions and filopodia of migrating cells. Mechanistically, the interaction depended on phosphorylated Fascin, active PKCgamma and active Rac, but not on active Cdc42. The activity of Rac on the Fascin/PKC complex was mediated in part by Pak. Elucidation of this novel pathway for regulation of the Fascin/PKCgamma complex in migrating carcinoma cells suggests novel targets for therapeutic intervention in metastasis.

  • dual actin bundling and protein kinase c binding activities of Fascin regulate carcinoma cell migration downstream of rac and contribute to metastasis
    Molecular Biology of the Cell, 2007
    Co-Authors: Yosuke Hashimoto, Maddy Parsons, Josephine C. Adams
    Abstract:

    Recurrence of carcinomas due to cells that migrate away from the primary tumor is a major problem in cancer treatment. Immunohistochemical analyses of human carcinomas have consistently correlated up-regulation of the actin-bundling protein Fascin with a clinically aggressive phenotype and poor prognosis. To understand the functional and mechanistic contributions of Fascin, we undertook inducible short hairpin RNA (shRNA) knockdown of Fascin in human colon carcinoma cells derived from an aggressive primary tumor. Fascin-depletion led to decreased numbers of filopodia and altered morphology of cell protrusions, decreased Rac-dependent migration on laminin, decreased turnover of focal adhesions, and, in vivo, decreased xenograft tumor development and metastasis. cDNA rescue of Fascin shRNA-knockdown cells with wild-type green fluorescent protein-Fascin or Fascins mutated at the protein kinase C (PKC) phosphorylation site revealed that both the actin-bundling and active PKC-binding activities of Fascin are required for the organization of filopodial protrusions, Rac-dependent migration, and tumor metastasis. Thus, Fascin contributes to carcinoma migration and metastasis through dual pathways that impact on multiple subcellular structures needed for cell migration.

  • interaction of Fascin and protein kinase cα a novel intersection in cell adhesion and motility
    The EMBO Journal, 2003
    Co-Authors: N. Anilkumar, Josephine C. Adams, Maddy Parsons, R D Monk
    Abstract:

    Coordination of protrusive and contractile cell–matrix contacts is important for cell adhesion and migration, but the mechanisms involved are not well understood. We report an unexpected direct association between Fascin, an actin-bundling component of filopodia, microspikes and lamellipodial ribs, and protein kinase Cα (PKCα), a regulator of focal adhesions. The association is detectable by protein–protein binding in vitro, by coimmunoprecipitation from cell extracts, and in live cells as fluorescence resonance energy transfer detected by fluorescence imaging lifetime microscopy. The interaction is physiologically regulated by the extracellular matrix context of cells, depends on activation of PKCα and is mediated by the C1B domain of PKCα. Strikingly, a Fascin mutant, Fascin S39D, associates constitutively with PKCα. Through use of a newly developed set of membrane-permeable peptides that separately inhibit either Fascin/PKCα or Fascin/actin binding, we have uncovered that specific blockade of the Fascin/PKCα interaction increases cell migration on fibronectin in conjunction with increased Fascin protrusions and remodeling of focal adhesions. These results identify the Fascin–PKCα interaction as an important novel intersection in the regulation and networking of cell–matrix contacts.

  • Fascins, and their roles in cell structure and function
    BioEssays : news and reviews in molecular cellular and developmental biology, 2002
    Co-Authors: Nina Kureishy, Vasileia Sapountzi, Soren Prag, N. Anilkumar, Josephine C. Adams
    Abstract:

    The Fascins are a structurally unique and evolutionarily conserved group of actin cross-linking proteins. Fascins function in the organisation of two major forms of actin-based structures: dynamic, cortical cell protrusions and cytoplasmic microfilament bundles. The cortical structures, which include filopodia, spikes, lamellipodial ribs, oocyte microvilli and the dendrites of dendritic cells, have roles in cell-matrix adhesion, cell interactions and cell migration, whereas the cytoplasmic actin bundles appear to participate in cell architecture. We discuss the current understanding of the cellular mechanisms that regulate the binding of Fascin to actin and how these processes contribute to the organisation or disassembly of cell protrusions. Although the in vivo roles of Fascin have been studied principally in Drosophila, several human diseases are associated with inherited or acquired alterations in the expression of Fascins. Strategies to modulate Fascin-containing protrusions and thereby cell adhesive and migratory behaviour could have potential for therapeutic intervention in these conditions. The supplementary material referred to in this section can be found at http://www.interscience.wiley.com/jpages/0265-9247/suppmat/2002/v24.350.html

Maddy Parsons - One of the best experts on this subject based on the ideXlab platform.

  • FMNL2 regulates dynamics of Fascin in filopodia.
    The Journal of cell biology, 2020
    Co-Authors: Karin Pfisterer, James A. Levitt, Campbell D Lawson, Richard J. Marsh, John M. Heddleston, Eric Wait, Simon Ameer-beg, Susan Cox, Maddy Parsons
    Abstract:

    Filopodia are peripheral F-actin-rich structures that enable cell sensing of the microenvironment. Fascin is an F-actin-bundling protein that plays a key role in stabilizing filopodia to support efficient adhesion and migration. Fascin is also highly up-regulated in human cancers, where it increases invasive cell behavior and correlates with poor patient prognosis. Previous studies have shown that Fascin phosphorylation can regulate F-actin bundling, and that this modification can contribute to subcellular Fascin localization and function. However, the factors that regulate Fascin dynamics within filopodia remain poorly understood. In the current study, we used advanced live-cell imaging techniques and a Fascin biosensor to demonstrate that Fascin phosphorylation, localization, and binding to F-actin are highly dynamic and dependent on local cytoskeletal architecture in cells in both 2D and 3D environments. Fascin dynamics within filopodia are under the control of formins, and in particular FMNL2, that binds directly to dephosphorylated Fascin. Our data provide new insight into control of Fascin dynamics at the nanoscale and into the mechanisms governing rapid cytoskeletal adaptation to environmental changes. This filopodia-driven exploration stage may represent an essential regulatory step in the transition from static to migrating cancer cells.

  • Fascin promotes filopodia formation independent of its role in actin bundling
    The Journal of cell biology, 2012
    Co-Authors: Jennifer Zanet, Maddy Parsons, Asier Jayo, Serge Plaza, Thomas H. Millard, Brian Stramer
    Abstract:

    Fascin is an evolutionarily conserved actin-binding protein that plays a key role in forming filopodia. It is widely thought that this function involves Fascin directly bundling actin filaments, which is controlled by an N-terminal regulatory serine residue. In this paper, by studying cellular processes in Drosophila melanogaster that require Fascin activity, we identify a regulatory residue within the C-terminal region of the protein (S289). Unexpectedly, although mutation (S289A) of this residue disrupted the actin-bundling capacity of Fascin, Fascin S289A fully rescued filopodia formation in Fascin mutant flies. Live imaging of migrating macrophages in vivo revealed that this mutation restricted the localization of Fascin to the distal ends of filopodia. The corresponding mutation of human Fascin (S274) similarly affected its interaction with actin and altered filopodia dynamics within carcinoma cells. These data reveal an evolutionarily conserved role for this regulatory region and unveil a function for Fascin, uncoupled from actin bundling, at the distal end of filopodia.

  • Rac regulates the interaction of Fascin with protein kinase C in cell migration.
    Journal of Cell Science, 2008
    Co-Authors: Maddy Parsons, Josephine C. Adams
    Abstract:

    Fascin is an actin-bundling protein that is low or absent in normal epithelia; its upregulation correlates with poor prognosis in many human carcinomas. We have recently demonstrated in mouse xenograft models that Fascin contributes to tumour development and metastasis through its dual actin-bundling and active PKC-binding activities. Rac was implicated as a regulator of Fascin-dependent colon carcinoma cell migration in vitro. Here, we tested the hypothesis that Rac regulates the interaction of Fascin with active PKC. The major conventional PKC in colon carcinoma cells is protein kinase Cgamma (PKCgamma). Endogenous PKCgamma, Fascin and Rac1 colocalised at lamellipodial margins of migrating cells. Colocalisation of Fascin and PKCgamma depended on Rac activity, and inhibition of Rac decreased PKCgamma activity in cell extracts but not in vitro. Fluorescence resonance energy transfer/fluorescence lifetime imaging microscopy uncovered that Fascin and PKCgamma interact in protrusions and filopodia of migrating cells. Mechanistically, the interaction depended on phosphorylated Fascin, active PKCgamma and active Rac, but not on active Cdc42. The activity of Rac on the Fascin/PKC complex was mediated in part by Pak. Elucidation of this novel pathway for regulation of the Fascin/PKCgamma complex in migrating carcinoma cells suggests novel targets for therapeutic intervention in metastasis.

  • dual actin bundling and protein kinase c binding activities of Fascin regulate carcinoma cell migration downstream of rac and contribute to metastasis
    Molecular Biology of the Cell, 2007
    Co-Authors: Yosuke Hashimoto, Maddy Parsons, Josephine C. Adams
    Abstract:

    Recurrence of carcinomas due to cells that migrate away from the primary tumor is a major problem in cancer treatment. Immunohistochemical analyses of human carcinomas have consistently correlated up-regulation of the actin-bundling protein Fascin with a clinically aggressive phenotype and poor prognosis. To understand the functional and mechanistic contributions of Fascin, we undertook inducible short hairpin RNA (shRNA) knockdown of Fascin in human colon carcinoma cells derived from an aggressive primary tumor. Fascin-depletion led to decreased numbers of filopodia and altered morphology of cell protrusions, decreased Rac-dependent migration on laminin, decreased turnover of focal adhesions, and, in vivo, decreased xenograft tumor development and metastasis. cDNA rescue of Fascin shRNA-knockdown cells with wild-type green fluorescent protein-Fascin or Fascins mutated at the protein kinase C (PKC) phosphorylation site revealed that both the actin-bundling and active PKC-binding activities of Fascin are required for the organization of filopodial protrusions, Rac-dependent migration, and tumor metastasis. Thus, Fascin contributes to carcinoma migration and metastasis through dual pathways that impact on multiple subcellular structures needed for cell migration.

  • interaction of Fascin and protein kinase cα a novel intersection in cell adhesion and motility
    The EMBO Journal, 2003
    Co-Authors: N. Anilkumar, Josephine C. Adams, Maddy Parsons, R D Monk
    Abstract:

    Coordination of protrusive and contractile cell–matrix contacts is important for cell adhesion and migration, but the mechanisms involved are not well understood. We report an unexpected direct association between Fascin, an actin-bundling component of filopodia, microspikes and lamellipodial ribs, and protein kinase Cα (PKCα), a regulator of focal adhesions. The association is detectable by protein–protein binding in vitro, by coimmunoprecipitation from cell extracts, and in live cells as fluorescence resonance energy transfer detected by fluorescence imaging lifetime microscopy. The interaction is physiologically regulated by the extracellular matrix context of cells, depends on activation of PKCα and is mediated by the C1B domain of PKCα. Strikingly, a Fascin mutant, Fascin S39D, associates constitutively with PKCα. Through use of a newly developed set of membrane-permeable peptides that separately inhibit either Fascin/PKCα or Fascin/actin binding, we have uncovered that specific blockade of the Fascin/PKCα interaction increases cell migration on fibronectin in conjunction with increased Fascin protrusions and remodeling of focal adhesions. These results identify the Fascin–PKCα interaction as an important novel intersection in the regulation and networking of cell–matrix contacts.

Zhong-ying Shen - One of the best experts on this subject based on the ideXlab platform.

  • Phosphorylation of Fascin decreases the risk of poor survival in patients with esophageal squamous cell carcinoma.
    The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2010
    Co-Authors: Qing Zhao, Jin-hui Shen, Zhong-ying Shen, Jian-jun Xie, Qiao Huang
    Abstract:

    Phosphorylation of Fascin at serine 39 (phospho-S39-Fascin) could inhibit its actin-binding and actin-bundling activities and decrease filopodia formation. However, the relationship between phospho-S39-Fascin expression and clinicopathological parameters in tumors is still unknown. Here, Western blot analysis and IHC applied to tissue microarray technology were performed to examine the expression status of non-phosphorylated Fascin (Fascin) and phospho-S39-Fascin and their impacts on the prognosis of patients with esophageal squamous cell carcinoma (ESCC). Fascin and phospho-S39-Fascin expressions were tested by cytoplasmic staining. Among the 254 patients, 90 cases showed high expression of Fascin and 87 cases showed high expression of phospho-S39-Fascin. Survival analysis showed that high expression of Fascin was significantly associated with a poor prognosis of the patients with ESCC (p=0.004). In contrast, high expression of phospho-S39-Fascin correlated significantly with an improved outcome of patients (p=0.020). Multivariate analysis showed that both Fascin and phospho-S39-Fascin were independent prognostic factors. In a combined analysis, the patients with high expression of Fascin and low expression of phospho-S39-Fascin tumors had a shorter overall survival than those with high expression of both Fascin and phospho-S39-Fascin tumors (5-year overall survival rate: 28.7% vs 48.3%, p=0.068). Our results suggest that high expression of Fascin correlates with poor outcome and that high expression of phospho-S39-Fascin decreases the risk of poor prognosis in ESCC. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.

  • involvement of cyr61 and ctgf in the Fascin mediated proliferation and invasiveness of esophageal squamous cell carcinomas cells
    American Journal of Pathology, 2010
    Co-Authors: Jian-jun Xie, Qing Zhao, Zhong-ying Shen, Feng Pan, Dong Xie
    Abstract:

    Fascin is over-expressed in esophageal squamous cell carcinoma (ESCC) and involved in the proliferation and invasiveness of ESCC cells. In this study, we retrospectively examined the expression of Fascin in ESCC samples by immunohistochemistry and revealed that overexpression of Fascin was related to poor patient survival. RNAi-mediated knockdown of Fascin in ESCC cells significantly inhibited cell proliferation and invasiveness, whereas forced expression of Fascin in immortalized esophageal epithelial cells accelerated cell proliferation and invasiveness. To explore the underlying mechanism, cDNA microarray was performed to identify the differential gene expression profiles between a Fascin-depleted cell line by RNAi and the corresponding control ESCC cells. Results showed that 296 genes were differentially expressed on Fascin depletion. In this study, we focused on two down-regulated genes: CYR61 and CTGF. We found that restored expression of either CYR61 or CTGF led to a recovery of the suppression of cellular proliferation and invasiveness induced by down-regulation of Fascin expression; the protein level of CYR61 and CTGF were up-regulated in ESCCs and their expression pattern correlated with Fascin overexpression. Finally, analysis of signal transduction revealed that Fascin affected the expressions of CYR61 and CTGF through transforming growth factor (TGF)-β pathway. Taken together, we propose that Fascin regulates the proliferation and invasiveness of ESCC cells by modulating the expression of CTGF and CYR61 via TGF-β pathway.

  • Fascin Expression in Human Embryonic, Fetal, and Normal Adult Tissue
    The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2007
    Co-Authors: Fa-ren Zhang, Li-hua Tao, Zhong-ying Shen
    Abstract:

    This study investigates the distribution of Fascin in human embryonic, fetal, and normal adult tissues. Tissue microarray technology was used to perform immunohistochemical experiments on human embryos and fetuses at 4–22 weeks of gestation and adult specimens. Fascin was widely expressed in the nervous system. At 4 weeks of gestation, Fascin was present in the neural tube. At 8–12 weeks of gestation, homogenous gene expression was seen in cells of the cerebellum and gastrointestinal tract. In later developmental stages and in adults, Purkinje cells of the cerebellum and glandular epithelium of the gastrointestinal tract showed no expression. Fascin was expressed in the cortex and medulla of the adrenal gland at 8–12 weeks of gestation, whereas immunoreactivity decreased from the zona glomerulosa through the zona reticularis and was essentially negative in the adrenal medulla of adults. Significant expression of Fascin was seen throughout development in neurons, follicular dendritic cells of lymphoid tissue, basal layer cells of stratified squamous epithelia, mesenchyme, and vascular endothelial cells. Simple columnar epithelia of the biliary duct, colon, ovary, pancreas, and stomach were all negative for Fascin expression. These results show that expression of Fascin is time specific and highly tissue specific. Parallels between Fascin expression in embryogenesis and carcinogenesis are discussed. (J Histochem Cytochem 56:193–199, 2008)

  • Fascin is a potential biomarker for early-stage oesophageal squamous cell carcinoma
    Journal of clinical pathology, 2006
    Co-Authors: Haihua Zhang, Jian-jun Xie, Wei-jia Cai, Dawei Xiao, Hongmei Zeng, Yongdong Niu, Zhangmin Yang, Zhong-ying Shen
    Abstract:

    Background: Fascin, an actin-binding protein, is usually expressed at a low level in normal epithelium, but is markedly up regulated in several types of carcinomas. Reports on Fascin expression in oesophageal squamous cell carcinoma (ESCC) and precancerous lesions remain rare. Aim: To show the roles of Fascin in the progression from normal epithelium to invasive ESCC. Methods: Fascin expression in 102 sections embedded in paraffin wax, including samples of normal mucosa (n = 20), dysplasia (n = 10), ESCC (n = 62) and special sections (n = 10) of a full-length mucosa layer from the distant margin to the cancer focus of the excised oesophagus, and 49 fresh specimens of ESCC was analysed by immunohistochemistry, western blot and real-time reverse transcription-polymerase chain reaction. Fascin expression in ESCC cell lines was also investigated. Results: In the immunohistochemical study, the positive rate of Fascin was significantly higher in the tumour tissue than in the normal epithelium (p = 0.020), but no significant difference was shown between ESCC and dysplasia (p = 1.000). Immunostaining for Fascin was only apparent in the basal layer of the normal epithelium. However, in the dysplasia, positive staining was observed in most of the heterogeneous cells from the basal layer to the granular layer of the epithelium. Fascin expression was seen to increase progressively from the normal epithelium to invasive ESCC. Up regulation of Fascin was observed in 87.76% (43/49) and 77.55% (38/49) of the specimens, respectively, using western blot and real-time reverse transcription-polymerase chain reaction assays; 80% (4/5) of ESCC cell lines also expressed Fascin at a high level. Furthermore, overexpression of Fascin was markedly correlated with cell proliferation and lymph node metastasis. Conclusions: These findings suggested that Fascin was associated with the transformation and development of ESCC and implicated the potential of Fascin as a novel biomarker that would allow the tumour to be identified at an early stage in high-risk individuals.

  • Role of Fascin in the proliferation and invasiveness of esophageal carcinoma cells
    Biochemical and biophysical research communications, 2005
    Co-Authors: Jian-jun Xie, Haihua Zhang, Wei-jia Cai, R.q. Mai, Yang-min Xie, Z.m. Yang, Y.d. Niu, Zhong-ying Shen
    Abstract:

    Fascin, an actin-bundling protein, induces membrane protrusions and increases cell motility in various transformed cells. The overexpression of Fascin in esophageal squamous cell carcinoma (ESCC) has been described only recently, but the roles and mechanism still remained unclear. Here, by using RNA interference (RNAi), we have stably silenced the expression of the Fascin in EC109 cells, an ESCC cell line. Down-regulation of Fascin resulted in a suppression of cell proliferation and as well as a decrease in cell invasiveness. Furthermore, we revealed that Fascin might have functions in regulating tumor growth in vivo. The effect of Fascin on cell invasiveness correlated with the activation of matrix metalloproteases such as MMP-2 and MMP-9. We examined that Fascin down-expression also led to a decrease of c-erbB-2 and β-catenin at the protein level. These results suggested that Fascin might play crucial roles in regulating neoplasm progression of ESCC.

Jianwei Sun - One of the best experts on this subject based on the ideXlab platform.

  • Fascin induces melanoma tumorigenesis and stemness through regulating the Hippo pathway.
    Cell communication and signaling : CCS, 2018
    Co-Authors: Jiaxin Kang, Jian Wang, Zhuang Yao, Qin Fan, Feng Gao, Yan Sun, Jianwei Sun
    Abstract:

    Fascin is a F-actin bundling protein and its overexpression is correlated with poor prognosis and increases metastatic potential in a number of cancers. But underlying function and mechanism of Fascin on tumorigenesis in melanoma remain elusive. The melanoma cell lines WM793 and WM39 were employed for the soft agar and sphere formation assay. Quantitative RT-PCR and Western blot were performed for identifying the gene expression at mRNA and protein levels, respectively. Co-IP and in vitro GST pulldown experiments were used to test the interaction between Fascin and MST2. Fascin regulates tumorigenesis and cancer cell stemness in melanoma through inhibition of the Hippo pathway kinase MST2 and the activation of transcription factor TAZ. Our data showed that Fascin interacts with the kinase domain of MST2 to inhibit its homodimer formation and kinase activity. Depletion of Fascin led to increase of p-LATS level and decrease of TAZ, but not YAP. We also demonstrated that Fascin regulates melanoma tumorigenesis independent of its actin-bundling activity. Fascin is a new regulator of the MST2-LATS-TAZ pathway and plays a critical role in melanoma tumorigenesis. Inhibition of Fascin reduces melanoma tumorigenesis and stemness, and thus Fascin could be a potential therapeutic target for this malignancy.

  • Fascin induces melanoma tumorigenesis and stemness through regulating the Hippo pathway
    BMC, 2018
    Co-Authors: Jiaxin Kang, Jian Wang, Zhuang Yao, Qin Fan, Feng Gao, Yan Sun, Jianwei Sun
    Abstract:

    Abstract Background Fascin is a F-actin bundling protein and its overexpression is correlated with poor prognosis and increases metastatic potential in a number of cancers. But underlying function and mechanism of Fascin on tumorigenesis in melanoma remain elusive. Methods The melanoma cell lines WM793 and WM39 were employed for the soft agar and sphere formation assay. Quantitative RT-PCR and Western blot were performed for identifying the gene expression at mRNA and protein levels, respectively. Co-IP and in vitro GST pulldown experiments were used to test the interaction between Fascin and MST2. Results Fascin regulates tumorigenesis and cancer cell stemness in melanoma through inhibition of the Hippo pathway kinase MST2 and the activation of transcription factor TAZ. Our data showed that Fascin interacts with the kinase domain of MST2 to inhibit its homodimer formation and kinase activity. Depletion of Fascin led to increase of p-LATS level and decrease of TAZ, but not YAP. We also demonstrated that Fascin regulates melanoma tumorigenesis independent of its actin-bundling activity. Conclusions Fascin is a new regulator of the MST2-LATS-TAZ pathway and plays a critical role in melanoma tumorigenesis. Inhibition of Fascin reduces melanoma tumorigenesis and stemness, and thus Fascin could be a potential therapeutic target for this malignancy

  • Monoubiquitination Inhibits the Actin Bundling Activity of Fascin.
    The Journal of biological chemistry, 2016
    Co-Authors: Shengchen Lin, Jianwei Sun, Jihui Hao, Mentor Mulaj, Bin Fang, Tyler Keeley, Lixin Wan, Martin Muschol, Shengyu Yang
    Abstract:

    Fascin is an actin bundling protein that cross-links individual actin filaments into straight, compact, and stiff bundles, which are crucial for the formation of filopodia, stereocillia, and other finger-like membrane protrusions. The dysregulation of Fascin has been implicated in cancer metastasis, hearing loss, and blindness. Here we identified monoubiquitination as a novel mechanism that regulates Fascin bundling activity and dynamics. The monoubiquitination sites were identified to be Lys247 and Lys250, two residues located in a positive charge patch at the actin binding site 2 of Fascin. Using a chemical ubiquitination method, we synthesized chemically monoubiquitinated Fascin and determined the effects of monoubiquitination on Fascin bundling activity and dynamics. Our data demonstrated that monoubiquitination decreased the Fascin bundling EC50, delayed the initiation of bundle assembly, and accelerated the disassembly of existing bundles. By analyzing the electrostatic properties on the solvent-accessible surface of Fascin, we proposed that monoubiquitination introduced steric hindrance to interfere with the interaction between actin filaments and the positively charged patch at actin binding site 2. We also identified Smurf1 as a E3 ligase regulating the monoubiquitination of Fascin. Our findings revealed a previously unidentified regulatory mechanism for Fascin, which will have important implications for the understanding of actin bundle regulation under physiological and pathological conditions.

  • hypoxia inducible factor 1 promotes pancreatic ductal adenocarcinoma invasion and metastasis by activating transcription of the actin bundling protein Fascin
    Cancer Research, 2014
    Co-Authors: Xiao Zhao, Song Gao, He Ren, Wei Sun, Huan Zhang, Jianwei Sun, Shengyu Yang, Jihui Hao
    Abstract:

    Because of the early onset of local invasion and distant metastasis, pancreatic ductal adenocarcinoma (PDAC) is the most lethal human malignant tumor, with a 5-year survival rate of less than 5%. In this study, we investigated the role of Fascin, a prometastasis actin-bundling protein, in PDAC progression, invasion, and the molecular mechanisms underlying Fascin overexpression in PDAC. Our data showed that the expression levels of Fascin were higher in cancer tissues than in normal tissues, and Fascin overexpression correlated with the PDAC differentiation and prognosis. Fascin overexpression promoted PDAC cell migration and invasion by elevating matrix metalloproteinase-2 (MMP-2) expression. Fascin regulated MMP-2 expression through protein kinase C and extracellular signal-regulated kinase. Importantly, our data showed that hypoxia induced Fascin overexpression in PDAC cells by promoting the binding of hypoxia-inducible factor-1 (HIF-1) to a hypoxia response element on the Fascin promoter and transactivating Fascin mRNA transcription. Intriguingly, HIF-1α expression levels in PDAC patient specimens significantly correlated with Fascin expression. Moreover, immunohistochemistry staining of consecutive sections demonstrated colocalization between HIF-1α and Fascin in PDAC specimens, suggesting that hypoxia and HIF-1α were responsible for Fascin overexpression in PDAC. When ectopically expressed, Fascin was able to rescue PDAC cell invasion after HIF-1α knockdown. Our results demonstrated that Fascin is a direct target gene of HIF-1. Our data suggested that the hypoxic tumor microenvironment in PDAC might promote invasion and metastasis by inducing Fascin overexpression, and Fascin might be targeted to block PDAC progression.

  • Fascin protein is critical for transforming growth factor β protein induced invasion and filopodia formation in spindle shaped tumor cells
    Journal of Biological Chemistry, 2011
    Co-Authors: Jianwei Sun, Yin Xiong, Junling Shen, Anna Cheng, Wei Chiao Chang, Mingfeng Hou, Johnathan M Lancaster, Minjung Kim, Shengyu Yang
    Abstract:

    Fascin, an actin-bundling protein overexpressed in all carcinomas, has been associated with poor prognosis, shorter survival, and more metastatic diseases. It is believed that Fascin facilitates tumor metastasis by promoting the formation of invasive membrane protrusions. However, the mechanisms by which Fascin is overexpressed in tumors are not clear. TGFβ is a cytokine secreted by tumor and mesenchymal cells and promotes metastasis in many late stage tumors. The pro-metastasis mechanisms of TGFβ remain to be fully elucidated. Here we demonstrated that TGFβ induced Fascin expression in spindle-shaped tumor cells through the canonical Smad-dependent pathway. Fascin was critical for TGFβ-promoted filopodia formation, migration, and invasion in spindle tumor cells. More importantly, Fascin expression significantly correlates with TGFβ1 and TGFβ receptor I levels in a cohort of primary breast tumor samples. Our results indicate that elevated TGFβ level in the tumor microenvironment may be responsible for Fascin overexpression in some of the metastatic tumors. Our data also suggest that Fascin could play a central role in TGFβ-promoted tumor metastasis.

Tarek A Bismar - One of the best experts on this subject based on the ideXlab platform.

  • Fascin regulates prostate cancer cell invasion and is associated with metastasis and biochemical failure in prostate cancer
    Clinical Cancer Research, 2009
    Co-Authors: Andrew Darnel, Emy Behmoaram, Robin T Vollmer, Jacques Corcos, Krikor Bijian, Kanishka Sircar, Jinsong Jiao, Moulaya A Alaouijamali, Tarek A Bismar
    Abstract:

    Purpose: Prostate cancer metastasis to secondary organs is considered an initial event in the development of hormone refractory disease and remains the major cause of death among prostate cancer patients. In this study, we investigated the role of Fascin, a cytoskeleton actin–bundling protein involved in the formation of filopodia and cell migration, in prostate cancer progression. Experimental Design: Fascin protein expression was examined by immunohistochemistry in a cohort of 196 patients with localized prostate cancer and across several stages of disease progression, including hormone refractory disease. Cellular changes were also assessed in vitro and in vivo in DU145 prostate cancer cell line using Fascin gene silencing. Results: Fascin epithelial expression was significantly up-regulated in localized and hormone refractory prostate cancer compared with benign prostate tissue ( P P = 0.075), signifying more aggressive clinical course, thus supporting a function for Fascin in prostate cancer progression. In cellular models, Fascin gene silencing using small interfering RNA in the androgen-independent prostate cancer cell line DU145 decreased cell motility and invasiveness while increasing cell adhesive properties. In addition, Fascin small interfering RNA–expressing DU145 cells implanted orthotopically in mouse prostate showed significantly decreased growth ( P P Conclusions: Our data show a function of Fascin in the regulation of prostate cancer progression and emphasize the importance of Fascin as a prognostic marker for aggressive disease and as a potential therapeutic target for advanced androgen independent disease.