DOCSLIB.ORG

Sonic Hedgehog/Gli1 Signaling Pathway Regulates Cell Migration and Invasion Via Induction of Epithelial-To

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Sonic Hedgehog/Gli1 Signaling Pathway Regulates Cell Migration and Invasion Via Induction of Epithelial-To Journal of Cancer 2020, Vol. 11 3932 Ivyspring International Publisher Journal of Cancer 2020; 11(13): 3932-3943. doi: 10.7150/jca.42900 Research Paper Sonic Hedgehog/Gli1 Signaling Pathway Regulates Cell Migration and Invasion via Induction of Epithelial-to- mesenchymal Transition in Gastric Cancer Bin Ke, Xiao-Na Wang, Ning Liu, Bin Li, Xue-Jun Wang, Ru-Peng Zhang, Han Liang Department of Gastric Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin’s Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy of Tianjin, Tianjin, 300060, P.R. China Corresponding author: Han Liang, Department of Gastric Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin’s Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy of Tianjin, Huan-Hu-Xi Road, HeXi District, Tianjin, 300060, People’s Republic of China. E-mail: [email protected] © The author(s). This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions. Received: 2019.12.09; Accepted: 2020.03.23; Published: 2020.04.06 Abstract Background: The aberrant activation of the Sonic hedgehog (Shh) signaling pathway is involved in progression of several types of cancer, including gastric cancer (GC). However, it remains uncertain whether it also plays a critical role in promoting cancer initiation and progression by inducing epithelial-to-mesenchymal transition (EMT) in GC. Thus, the aim of the present study was to determine whether the Shh pathway is involved in GC, and to investigate the function of the Shh pathway in the induction of EMT in GC. Materials and methods: The expression levels of Shh pathway members and EMT markers were examined in GC tissues by immunohistochemistry. The association between these factors and patient clinicopathological characteristics was analyzed. In addition, Gli-antagonist 61 (GANT61) was used to block Shh/Gli1 pathway activity, and recombinant Shh proteins (N-Shh) were used to activate the Shh pathway in GC cells. Wound healing and Transwell invasion and migration assays were performed to assess the effects of the Shh pathway on the migration and invasion of GC cells in vitro. Furthermore, western blot analysis was used to examine the changes in protein expression. Results: The results demonstrated that these Shh/Gli1 pathway members were upregulated in GC tissues, and that Gli1 upregulation was associated with tumor progression and a poor prognosis. Gli1 expression was negatively associated with E-cadherin (E-Cad) expression, and positively with Vimentin (VIM) expression in GC specimens. Further analysis revealed that when the Shh/Gli1 pathway was activated, the migratory and invasive abilities of GC cells were enhanced, and the expression levels of Gli1 and VIM were increased, while E-Cad expression was decreased. Opposite results were observed when the Shh/Gli1 pathway was blocked by GANT61. Conclusions: The present study indicated that the Shh/Gli1 pathway exhibits an abnormal activation pattern in GC with possible predictive and prognostic significance. The Shh/Gli1 pathway may promote the migratory and invasive potential of GC cells by inducing EMT. The Shh/Gli1 pathway can thus be considered as a potential therapeutic target for GC. Key words: sonic hedgehog, epithelial-mesenchymal transition, gastric cancer, invasion, migration Introduction Gastric cancer (GC) is the fifth most frequent approximately 783 thousand deaths reported type of cancer and the third primary cause of annually [1]. Despite recent improvements in cancer-associated mortality worldwide with diagnosis and therapy, the prognosis of patients with http://www.jcancer.org Journal of Cancer 2020, Vol. 11 3933 advanced-stage GC remains dismal, with the 5-year of invasion and metastasis in GC, and may aid the overall survival rate being <30% [2, 3]. Local tumor development of effective approaches for the invasion and distant metastasis may be the major prevention and therapy of this malignancy. reasons for the poor survival of patients with Recently, several studies have demonstrated that advanced GC [4]. Therefore, understanding the exact the abnormal activation of the Shh pathway affects molecular mechanisms underlying the development EMT in several types of cancers, such as and progression of GC is important for the hepatocellular carcinoma and lung cancer [25, 26]. development of novel treatment targets for patients However, the association between the Shh pathway with GC. and EMT in GC has rarely been reported, and whether The hedgehog (Hh) signaling pathway is a or not the Shh pathway may promote cancer well-conserved pathway across various species that development and progression by inducing EMT in plays a critical role during embryonic development GC has not yet been investigated, at least to the best of and tissue morphogenesis in adults [5, 6]. In humans, our knowledge. In the present study, we analyzed the the Hh pathway is composed of Hh ligands, two expression of Shh pathway components and patient transmembrane protein receptors [Patched receptor clinicopathological features and prognosis in GC (Path) and Smoothened receptor (Smo)], and the tissues to evaluate whether the Shh pathway is five-zinc finger transcription factor Glis [7]. In the involved in GC. In addition, the activation of the Shh canonical Hh pathway, when the Hh ligand is absent, pathway was regulated by treatment with exogenous Ptch suppresses the activity of Smo, and thus inhibits recombinant Shh protein (N-Shh) or Gli-antagonist 61 the cleavage of Gli, blocking Hh pathway activation. (GANT61), a Gli specific inhibitor, to further explore When the Hh ligand is present, it binds to Ptch, the detailed mechanisms of the role of the Shh relieving the inhibition of Smo and resulting in the pathway in the regulation of EMT in GC. This study activation of Gli transcription factor. Activated Gli may indicate the potential clinical significance of the translocates to the nucleus and subsequently regulates Shh pathway and EMT in GC. the transcription of Gli target genes [8]. In mammals, three ligands have been identified in the Hh pathway: Materials and methods Sonic hedgehog (Shh), Desert hedgehog (Dhh) and Patients and specimens Indian hedgehog (Ihh) [7]. Among these three ligands, Shh is the most thoroughly studied one, as well as the A total of 178 GC specimens and 89 adjacent most important one [9, 10]. Recently, the ectopic non-tumor tissues were collected from patients activation of the Shh pathway has been detected in a diagnosed with GC at TianJin Medical University number of human malignancies [11, 12]. Numerous Cancer Institute and Hospital (TianJin, China) reports have indicated that the Shh pathway plays a between March 2010 and April 2013. All the patients crucial role in GC initiation, progression and underwent radical gastrectomy with D2 metastasis, and it has also been shown to be lymphadenectomy. None of the patients had associated with patient prognosis [13-15]. Therefore, a undergone chemotherapy, radiotherapy, or any other deeper understanding of this pathway in tumor treatment for cancer prior to surgery. The surgical invasion and metastasis may provide great promise tissues were formalin-fixed and paraffin-embedded. for GC therapy. The clinicopathological data were collected and Epithelial-to-mesenchymal transition (EMT) analyzed. The pathological tumor staging was exerts marked effects on cellular transdifferentiation classified according to the 8th edition of the American during embryonic development [16], and has been Joint Committee on Cancer TNM staging system for characterized as cells acquiring mesenchymal GC [27]. spindle-like morphology and the loosening of This study was approved by the Ethics intercellular cohesion [17, 18]. The characteristic Committee of TianJin Medical University Cancer features of EMT are the decreased expression of the Institute and Hospital (Tianjin, China). cell adhesion molecule, E-cadherin (E-Cad), and the Cell culture and treatment enhanced expression of the mesenchymal marker, The human gastric adenocarcinoma cell lines, Vimentin (VIM) [19, 20]. Previous studies have KATO, MKN-45, SGC-7901, MGC-803, AGS, N87 and demonstrated that EMT is a vital process in tumor BGC-823, and the human gastric epithelial cell line, progression and metastasis, and is associated with a GES-1, were obtained from the American Type poor prognosis and tumor aggressiveness in several Culture Collection (ATCC). The cell lines were human epithelial cancers, including GC [21-24]. The maintained in RPMI-1640 (Life Technologies; Thermo identification of factors that induce EMT would Fisher Scientific, Inc.) medium with 10% fetal bovine greatly contribute to understanding the mechanisms serum (FBS) (Thermo Fisher Scientific, Inc.), and 1% http://www.jcancer.org Journal of Cancer 2020, Vol. 11 3934 penicillin-streptomycin solution (Thermo Fisher branes (EMD Millipore). The membranes were Scientific, Inc.) at 37˚C in a humidified atmosphere of blocked with 5% skimmed milk for 1 h at room 5% CO2. temperature and then incubated with primary All cultures were routinely grown to ~70% antibodies at 4˚C overnight. After washing, the confluence one day before treatment. For the membranes were exposed to horseradish peroxidase inhibition of the Shh
Load more

Recommended publications
  • Hedgehog Signaling Is Evolutionarily Conserved Cilium-Independent
    Downloaded from genesdev.cshlp.org on August 14, 2009 - Published by Cold Spring Harbor Laboratory Press Cilium-independent regulation of Gli protein function by Sufu in Hedgehog signaling is evolutionarily conserved Miao-Hsueh Chen, Christopher W. Wilson, Ya-Jun Li, et al. Genes Dev. 2009 23: 1910-1928 Access the most recent version at doi:10.1101/gad.1794109 Supplemental http://genesdev.cshlp.org/content/suppl/2009/07/23/23.16.1910.DC1.html Material References This article cites 97 articles, 47 of which can be accessed free at: http://genesdev.cshlp.org/content/23/16/1910.full.html#ref-list-1 Email alerting Receive free email alerts when new articles cite this article - sign up in the box at the service top right corner of the article or click here To subscribe to Genes & Development go to: http://genesdev.cshlp.org/subscriptions Copyright © 2009 by Cold Spring Harbor Laboratory Press Downloaded from genesdev.cshlp.org on August 14, 2009 - Published by Cold Spring Harbor Laboratory Press Cilium-independent regulation of Gli protein function by Sufu in Hedgehog signaling is evolutionarily conserved Miao-Hsueh Chen,1,3 Christopher W. Wilson,1,3 Ya-Jun Li,1 Kelvin King Lo Law,2 Chi-Sheng Lu,1 Rhodora Gacayan,1 Xiaoyun Zhang,2 Chi-chung Hui,2 and Pao-Tien Chuang1,4 1Cardiovascular Research Institute, University of California at San Francisco, San Francisco, California 94158, USA; 2Program in Developmental and Stem Cell Biology, The Hospital for Sick Children, and Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5G 1L7, Canada A central question in Hedgehog (Hh) signaling is how evolutionarily conserved components of the pathway might use the primary cilium in mammals but not fly.
    [Show full text]
  • GLI1-Amplifications Expand the Spectrum of Soft Tissue Neoplasms Defined by GLI1 Gene Fusions
    Modern Pathology (2019) 32:1617–1626 https://doi.org/10.1038/s41379-019-0293-x ARTICLE GLI1-amplifications expand the spectrum of soft tissue neoplasms defined by GLI1 gene fusions 1 1 1 2 3 Narasimhan P. Agaram ● Lei Zhang ● Yun-Shao Sung ● Samuel Singer ● Todd Stevens ● 3 4 5 6 6 Carlos N. Prieto-Granada ● Justin A. Bishop ● Benjamin A. Wood ● David Swanson ● Brendan C. Dickson ● Cristina R. Antonescu1 Received: 18 February 2019 / Revised: 29 April 2019 / Accepted: 1 May 2019 / Published online: 12 June 2019 © United States & Canadian Academy of Pathology 2019 Abstract GLI1 fusions involving ACTB, MALAT1, and PTCH1 genes have been recently reported in a subset of malignant soft tissue tumors with characteristic monomorphic nested epithelioid morphology and frequent S100 positivity. However, we encountered a group of morphologically similar soft tissue tumors lacking the canonical GLI1 gene fusions and sought to investigate their genetic abnormalities. A combined approach including RNA sequencing, targeted exome sequencing and FISH methodologies were used to identify potential novel genetic abnormalities. Ten patients (five females, five males) with – fi 1234567890();,: 1234567890();,: an age range of 4 65 years (median 32.5) were identi ed. Tumors were located in the soft tissues of the limbs, trunk and head and neck, with one each in the tongue and lung. Histologically, tumors revealed ovoid to epithelioid cells arranged in a distinctive nested-trabecular pattern, separated by thin septa and a delicate vascular network. Two cases showed areas of increased nuclear pleomorphism and focal fascicular spindle cell growth. Four tumors showed a high mitotic count (≥15/10 HPFs), with necrosis seen in three of them.
    [Show full text]
  • Role of Sonic Hedgehog Signaling Pathway in Neuroblastoma Development
    Review Article Biology and Medicine, 1 (4): Rev2, 2009 eISSN: 09748369, www.biolmedonline.com Role of Sonic hedgehog signaling pathway in neuroblastoma development Mehdi Hayat Shahi1,2,3,§, Subrata Sinha2, *Mohammad Afzal3, *Javier S Castresana1 1Unidad de Biologia de Tumoures Cerebrales, Universidad de Navarra, 31008 Pamplona, Spain. 2Department of Biochemistry, All India Institute of Medical Sciences (AIIMS), New Delhi-110029, India. 3Section of Genetics, Department of Zoology, Aligarh Muslim University, Aligarh-202002, India. §Present address: Department of Genetics and Pathology, Rudbeck Laboratory, University of Uppsala, Uppsala- 75185, Sweden. *Corresponding Authors: Javier S Castresana, [email protected] Mohammad Afzal, [email protected] Abstract Malignant transformation of normal cells is a complex and accumulative process. Understanding this event gives insight into mechanisms of developmental biology and physical interaction of cellular machinery with surrounding ambient factors. However, the trend of embryonic malignancy is not interactive with ambient factors, rather a cause of deregulations of internal developmental process. In this review, we have attempted to explore the possibility of Sonic hedgehog role (Shh) in the development of neuroblastoma tumour. It is the major extra cranial tumour and develops in very early stage of childhood. Sonic hedgehog signaling is very well studied in another major childhood tumour i.e. medulloblastoma that contributes 20-25% of childhood tumours, and one-fourth of medulloblastoma is due to abnormality in the Shh signaling pathway. Therefore, we would consider whether Shh could also contribute to the development of neuroblastoma. Although scientists are coming up with the role of Shh in the neuroblastoma, the Sonic hedgehog signaling is very much one of the promising pathways because of its multi-dimensional role not only in CNS development but also in organogenesis and other major tumour development.
    [Show full text]
  • Sonic Hedgehog Signaling Limits Atopic Dermatitis Via Gli2-Driven Immune Regulation
    Sonic Hedgehog signaling limits atopic dermatitis via Gli2-driven immune regulation Eleftheria Papaioannou, … , Ryan F. L. O’Shaughnessy, Tessa Crompton J Clin Invest. 2019. https://doi.org/10.1172/JCI125170. Research Article Immunology Inflammation Hedgehog (Hh) proteins regulate development and tissue homeostasis, but their role in atopic dermatitis (AD) remains unknown. We found that on induction of mouse AD, Sonic Hedgehog (Shh) expression in skin, and Hh pathway action in skin T cells were increased. Shh signaling reduced AD pathology and the levels of Shh expression determined disease severity. Hh-mediated transcription in skin T cells in AD-induced mice increased Treg populations and their suppressive function through increased active transforming growth factor–b (TGF-b) in Tregs signaling to skin T effector populations to reduce disease progression and pathology. RNA sequencing of skin CD4+ T cells from AD-induced mice demonstrated that Hh signaling increased expression of immunoregulatory genes and reduced expression of inflammatory and chemokine genes. Addition of recombinant Shh to cultures of naive human CD4+ T cells in iTreg culture conditions increased FOXP3 expression. Our findings establish an important role for Shh upregulation in preventing AD, by increased Gli-driven Treg cell–mediated immune suppression, paving the way for a potential new therapeutic strategy. Find the latest version: http://jci.me/125170/pdf The Journal of Clinical Investigation RESEARCH ARTICLE Sonic Hedgehog signaling limits atopic dermatitis via Gli2-driven immune regulation Eleftheria Papaioannou,1 Diana C. Yánez,1,2 Susan Ross,1 Ching-In Lau,1 Anisha Solanki,1 Mira Manilal Chawda,1 Alex Virasami,1 Ismael Ranz,3 Masahiro Ono,1,4 Ryan F.
    [Show full text]
  • Non-Canonical Activation of Hedgehog in Prostate Cancer Cells Mediated by the Interaction of Transcriptionally Active Androgen Receptor Proteins with Gli3
    Oncogene (2018) 37:2313–2325 https://doi.org/10.1038/s41388-017-0098-7 ARTICLE Non-canonical activation of hedgehog in prostate cancer cells mediated by the interaction of transcriptionally active androgen receptor proteins with Gli3 1 1,2 2 1 1 1 2,3 Na Li ● Sarah Truong ● Mannan Nouri ● Jackson Moore ● Nader Al Nakouzi ● Amy Anne Lubik ● Ralph Buttyan Received: 19 July 2017 / Revised: 18 October 2017 / Accepted: 29 November 2017 / Published online: 12 February 2018 © The Author(s) 2018. This article is published with open access Abstract Hedgehog (Hh) is an oncogenic signaling pathway that regulates the activity of Gli transcription factors. Canonical Hh is a Smoothened-(Smo-) driven process that alters the post-translational processing of Gli2/Gli3 proteins. Though evidence supports a role for Gli action in prostate cancer (PCa) cell growth and progression, there is little indication that Smo is involved. Here we describe a non-canonical means for activation of Gli transcription in PCa cells mediated by the binding of transcriptionally-active androgen receptors (ARs) to Gli3. Androgens stimulated reporter expression from a Gli-dependent promoter in a variety of AR + PCa cells and this activity was suppressed by an anti-androgen, Enz, or by AR knockdown. 1234567890();,: Androgens also upregulated expression of endogenous Gli-dependent genes. This activity was associated with increased intranuclear binding of Gli3 to AR that was antagonized by Enz. Fine mapping of the AR binding domain on Gli2 showed that AR recognizes the Gli protein processing domain (PPD) in the C-terminus. Mutations in the arginine-/serine repeat elements of the Gli2 PPD involved in phosphorylation and ubiquitinylation blocked the binding to AR.
    [Show full text]
  • The Adrenal Capsule Is a Signaling Center Controlling Cell Renewal and Zonation Through Rspo3
    Downloaded from genesdev.cshlp.org on September 24, 2021 - Published by Cold Spring Harbor Laboratory Press RESEARCH COMMUNICATION The permanent cortex is formed through recruitment of The adrenal capsule is a capsular cells in a process that involves SHH signaling signaling center controlling (King et al. 2009). By E17.5, steroidogenic cells have adopted specific expression profiles, with the outermost cell renewal and zonation cell layers (zona glomerulosa [ZG]) producing enzymes Rspo3 that are required for mineralocorticoid production (e.g., through CYP11B2), and deeper layers (zona fasciculata [ZF]) Valerie Vidal,1,2,3,9 Sonia Sacco,1,2,3,9 expressing genes involved in glucocorticoid synthesis Ana Sofia Rocha,1,2,3,8 Fabio da Silva,1,2,3 (Cyp11b1). In humans, but not rodents, a third layer (zona reticularis) can be distinguished that produces an- Clara Panzolini,1,2,3 Typhanie Dumontet,4,5 1,2,3 6 drogens and is located close to the medulla. Several lines Thi Mai Phuong Doan, Jingdong Shan, of evidence suggest that β-catenin plays an important Aleksandra Rak-Raszewska,6 Tom Bird,7 role in adrenal zonation and maintenance. Activation of Seppo Vainio,6 Antoine Martinez,4,5 the β-catenin pathway is restricted to the ZG (Kim et al. and Andreas Schedl1,2,3 2008; Walczak et al. 2014), and ectopic expression leads to the activation of ZG markers in ZF cells (Berthon 1Institute of Biology Valrose, Université de Nice-Sophia, F-06108 et al. 2010). Moreover, β-catenin seems to bind to and con- Nice, France; 2UMR1091, Institut National de la Santé et de la trol the expression of At1r, a gene specifically expressed Recherche Médicale, F-06108 Nice, France; 3CNRS, UMR7277, within the ZG (Berthon et al.
    [Show full text]
  • GLI2 but Not GLI1/GLI3 Plays a Central Role in the Induction of Malignant Phenotype of Gallbladder Cancer
    ONCOLOGY REPORTS 45: 997-1010, 2021 GLI2 but not GLI1/GLI3 plays a central role in the induction of malignant phenotype of gallbladder cancer SHU ICHIMIYA1, HIDEYA ONISHI1, SHINJIRO NAGAO1, SATOKO KOGA1, KUKIKO SAKIHAMA2, KAZUNORI NAKAYAMA1, AKIKO FUJIMURA3, YASUHIRO OYAMA4, AKIRA IMAIZUMI1, YOSHINAO ODA2 and MASAFUMI NAKAMURA4 Departments of 1Cancer Therapy and Research, 2Anatomical Pathology, 3Otorhinolaryngology and 4Surgery and Oncology, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812‑8582, Japan Received August 10, 2020; Accepted December 7, 2020 DOI: 10.3892/or.2021.7947 Abstract. We previously reported that Hedgehog (Hh) signal Introduction was enhanced in gallbladder cancer (GBC) and was involved in the induction of malignant phenotype of GBC. In recent Gallbladder cancer (GBC) is the seventh most common gastro- years, therapeutics that target Hh signaling have focused on intestinal carcinoma and accounts for 1.2% of all cancer cases molecules downstream of smoothened (SMO). The three tran- and 1.7% of all cancer-related deaths (1). GBC develops from scription factors in the Hh signal pathway, glioma-associated metaplasia to dysplasia to carcinoma in situ and then to invasive oncogene homolog 1 (GLI1), GLI2, and GLI3, function down- carcinoma over 5‑15 years (2). During this time, GBC exhibits stream of SMO, but their biological role in GBC remains few characteristic symptoms, and numerous cases have already unclear. In the present study, the biological significance of developed into locally advanced or metastasized cancer by the GLI1, GLI2, and GLI3 were analyzed with the aim of devel- time of diagnosis. Gemcitabine (GEM), cisplatin (CDDP), and oping novel treatments for GBC.
    [Show full text]
  • Distinct Activities of Gli1 and Gli2 in the Absence of Ift88 and the Primary Cilia
    Journal of Developmental Biology Article Distinct Activities of Gli1 and Gli2 in the Absence of Ift88 and the Primary Cilia Yuan Wang 1,2,†, Huiqing Zeng 1,† and Aimin Liu 1,* 1 Department of Biology, Eberly College of Sciences, Center for Cellular Dynamics, Huck Institute of Life Science, The Penn State University, University Park, PA 16802, USA; [email protected] (Y.W.); [email protected] (H.Z.) 2 Department of Occupational Health, School of Public Health, China Medical University, No.77 Puhe Road, Shenyang North New Area, Shenyang 110122, China * Correspondence: [email protected]; Tel.: +1-814-865-7043 † These authors contributed equally to this work. Received: 2 November 2018; Accepted: 16 February 2019; Published: 19 February 2019 Abstract: The primary cilia play essential roles in Hh-dependent Gli2 activation and Gli3 proteolytic processing in mammals. However, the roles of the cilia in Gli1 activation remain unresolved due to the loss of Gli1 transcription in cilia mutant embryos, and the inability to address this question by overexpression in cultured cells. Here, we address the roles of the cilia in Gli1 activation by expressing Gli1 from the Gli2 locus in mouse embryos. We find that the maximal activation of Gli1 depends on the cilia, but partial activation of Gli1 by Smo-mediated Hh signaling exists in the absence of the cilia. Combined with reduced Gli3 repressors, this partial activation of Gli1 leads to dorsal expansion of V3 interneuron and motor neuron domains in the absence of the cilia. Moreover, expressing Gli1 from the Gli2 locus in the presence of reduced Sufu has no recognizable impact on neural tube patterning, suggesting an imbalance between the dosages of Gli and Sufu does not explain the extra Gli1 activity.
    [Show full text]
  • Mechanisms Acting on Hedgehog-Gli Pathway and Their Therapeutic Potential
    From the Department of Biosciences and Nutrition Karolinska Institutet, Stockholm, Sweden MECHANISMS ACTING ON HEDGEHOG-GLI PATHWAY AND THEIR THERAPEUTIC POTENTIAL Ani Azatyan Stockholm 2020 All previously published papers were reproduced with permission from the publisher. Published by Karolinska Institutet. Printed by Universitetsservice US-AB © Ani Azatyan, 2020 ISBN 978-91-7831-880-3 Mechanisms acting on Hedgehog-GLI pathway and their therapeutic potential THESIS FOR DOCTORAL DEGREE (Ph.D.) By Ani Azatyan Principal Supervisor: Opponent: Prof. Peter Zaphiropoulos Prof. Matthias Lauth Karolinska Institutet Philipps-University Marburg, Germany Department of Biosciences and Nutrition Center for Tumor Biology and Immunology Co-supervisor(s): Examination Board: Prof. Karl Ekwall Doc. Ning Xu Landén Karolinska Institutet Karolinska Institutet Department of Biosciences and Nutrition Department of Medicine, Division of Dermatology Doc. Vladimir Bykov Prof. Sonia Lain Karolinska Institutet Karolinska Institutet Department of Oncology-Pathology Department of Microbiology, Tumor and Cell Biology Prof. Ann-Kristin Östlund Farrants Stockholm University Department of Molecular Biosciences “I am among those who think that science has great beauty” - Marie Curie Abstract Hedgehog signaling is crucial for diverse aspects of animal development, essential in regulating many cellular processes and is largely implicated in various forms of human cancer. However, many aspects of Hedgehog signaling are not completely understood. This thesis aims to contribute towards a better understanding of the mechanisms acting on Hedgehog-GLI signaling and explore their possible therapeutic potential. PAPER I. We demonstrate that the small molecule RITA, a p53 activator, downregulates Hedgehog signaling in human medulloblastoma and rhabdomyosarcoma cells via JNK kinase and irrespective of p53. In vitro RITA enhanced the anti-proliferative effects of the GLI antagonist GANT61.
    [Show full text]
  • The Role of Highly Selective Androgen Receptor (AR) Targeted
    P h a s e I I S t u d y o f I t r a c o n a z o l e i n B i o c h e m i c a l R e l a p s e Version 4.0: October 8, 2014 CC# 125513 CC# 125513: Hedgehog Inhibition as a Non-Castrating Approach to Hormone Sensitive Prostate Cancer: A Phase II Study of Itraconazole in Biochemical Relapse Investigational Agent: Itraconazole IND: IND Exempt (IND 116597) Protocol Version: 4.0 Version Date: October 8, 2014 Principal Investigator: Rahul Aggarwal, M.D., HS Assistant Clinical Professor Division of Hematology/Oncology, Department of Medicine University of California San Francisco 1600 Divisadero St. San Francisco, CA94115 [email protected] UCSF Co-Investigators: Charles J. Ryan, M.D., Eric Small, M.D., Professor of Medicine Professor of Medicine and Urology Lawrence Fong, M.D., Terence Friedlander, M.D., Professor in Residence Assistant Clinical Professor Amy Lin, M.D., Associate Clinical Professor Won Kim, M.D., Assistant Clinical Professor Statistician: Li Zhang, Ph.D, Biostatistics Core RevisionHistory October 8, 2014 Version 4.0 November 18, 2013 Version 3.0 January 28, 2013 Version 2.0 July 16, 2012 Version 1.0 Phase II - Itraconazole Page 1 of 79 P h a s e I I S t u d y o f I t r a c o n a z o l e i n B i o c h e m i c a l R e l a p s e Version 4.0: October 8, 2014 CC# 125513 Protocol Signature Page Protocol No.: 122513 Version # and Date: 4.0 - October 8, 2014 1.
    [Show full text]
  • Hedgehog Signalling in the Tumourigenesis and Metastasis of Osteosarcoma, and Its Potential Value in the Clinical Therapy Of
    Yao et al. Cell Death and Disease (2018) 9:701 DOI 10.1038/s41419-018-0647-1 Cell Death & Disease REVIEW ARTICLE Open Access Hedgehog signalling in the tumourigenesis and metastasis of osteosarcoma, and its potential value in the clinical therapy of osteosarcoma Zhihong Yao1,LeiHan1, Yongbin Chen2,FeiHe3,BinSun3, Santosh kamar1, Ya Zhang1,YihaoYang1, Cao Wang1 and Zuozhang Yang1 Abstract The Hedgehog (Hh) signalling pathway is involved in cell differentiation, growth and tissue polarity. This pathway is also involved in the progression and invasion of various human cancers. Osteosarcoma, a subtype of bone cancer, is commonly seen in children and adolescents. Typically, pulmonary osteosarcoma metastases are especially difficult to control. In the present paper, we summarise recent studies on the regulation of osteosarcoma progression and metastasis by downregulating Hh signalling. We also summarise the crosstalk between the Hh pathway and other cancer-related pathways in the tumourigenesis of various cancers. We further summarise and highlight the therapeutic value of potential inhibitors of Hh signalling in the clinical therapy of human cancers. 1234567890():,; 1234567890():,; Facts Open questions ● The Hh pathway regulates the progression of ● How does the Hh pathway regulate the tumourigenic osteosarcoma. progression and invasion of human osteosarcoma? ● The Hh pathway is involved in the metastasis of ● How does the Hh pathway interact with other osteosarcoma into other organs, such as the lungs. cancer-related pathways in the progression and ● The Hh pathway crosstalks with other cancer-related metastasis of cancers? pathways in the tumourigenesis of cancers. ● Could the Hh pathway be used as a target or ● The therapeutic value of the Hh pathway in the biomarker in clinical therapy for human clinical therapy of osteosarcoma is summarised.
    [Show full text]
  • Nuclear Factor-KB Contributes to Hedgehog Signaling Pathway Activation Through Sonic Hedgehog Induction in Pancreatic Cancer
    Research Article Nuclear Factor-KB Contributes to Hedgehog Signaling Pathway Activation through Sonic Hedgehog Induction in Pancreatic Cancer Hiroshi Nakashima,1 Masafumi Nakamura,1 Hiroshi Yamaguchi,2 Naoki Yamanaka,1 Takashi Akiyoshi,1 Kenichiro Koga,1 Koji Yamaguchi,3 Masazumi Tsuneyoshi,2 Masao Tanaka,3 and Mitsuo Katano1 Departments of 1Cancer Therapy and Research, 2Anatomic Pathology, and 3Surgery and Oncology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan Abstract The hedgehog (Hh) signaling pathway is crucial to growth and The hedgehog (Hh) signaling pathway, which functions as an patterning in a wide variety of tissues during embryonic organizer in embryonic development, is implicated in the development (3, 4). Of three Hh ligands, sonic Hh (Shh), Indian development of various tumors. In pancreatic cancer, pathway Hh (Ihh), and desert Hh (Dhh), Shh is reported to play an essential activation is reported to result from aberrant expression of role in the development of pancreatic cancer as well as pancreatic organogenesis (5, 6). Shh undergoes extensive post-translational the ligand, sonic Hh (Shh). However, the details of the mechanisms regulating Shh expression are not yet known. modifications to become biologically active (7). The signal peptide We hypothesized that nuclear factor-KB (NF-KB), a hallmark is cleaved from the precursor form of Shh to yield the 45-kDa transcription factor in inflammatory responses, contributes to full-length form. Further processing of this molecular form of Shh the overexpression of Shh in pancreatic cancer. In the present generates a 19-kDa amino and 26-kDa COOH-terminal peptides. study, we found a close positive correlation between NF-KB The 19-kDa NH2-terminal peptide undergoes COOH-terminal p65 and Shh expression in surgically resected pancreas esterification to a cholesterol molecule before being secreted into specimens, including specimens of chronic pancreatitis and the extracellular spaces where it mediates the physiologic actions pancreatic adenocarcinoma.
    [Show full text]