DOCSLIB.ORG

A Molecular Role for Lysyl Oxidase in Breast Cancer Invasion1

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
A Molecular Role for Lysyl Oxidase in Breast Cancer Invasion1 [CANCER RESEARCH 62, 4478–4483, August 1, 2002] A Molecular Role for Lysyl Oxidase in Breast Cancer Invasion1 Dawn A. Kirschmann,2 Elisabeth A. Seftor, Sheri F. T. Fong, Daniel R. C. Nieva, Colleen M. Sullivan, Elijah M. Edwards, Pascal Sommer, Katalin Csiszar, and Mary J. C. Hendrix Department of Anatomy and Cell Biology, Holden Comprehensive Cancer Center at The University of Iowa, The Roy J. and Lucille A. Carver College of Medicine, Iowa City, Iowa 52242-1109 [D. A. K., E. A. S., D. R. C. N., C. M. S., E. M. E., M. J. C. H.]; Institut de Biologie et Chimie des Prote´ines, Lyon Cedex 07, France [P. S.]; and Pacific Biomedical Research Center, University of Hawaii, Honolulu, Hawaii 96822 [S. F. T. F., K. C.] ABSTRACT LOX is a copper-dependent amine oxidase that initiates the cova- lent cross-linking of collagens and elastin in extracellular matrices We identified previously an up-regulation in lysyl oxidase (LOX) ex- (reviewed in Refs. 3, 4). It is secreted as a M 50,000 glycosylated pression, an extracellular matrix remodeling enzyme, in a highly invasive/ r proenzyme, which is proteolytically processed by procollagen C pro- metastatic human breast cancer cell line, MDA-MB-231, compared with MCF-7, a poorly invasive/nonmetastatic breast cancer cell line. In this teinase (bone morphogenic protein-1) into a mature, biologically study, we demonstrate that the mRNA expression of LOX and other LOX active Mr 32,000 form (3, 4). The formation of collagen/elastin family members [lysyl oxidase-like (LOXL), LOXL2, LOXL3, and cross-links by LOX leads to an increase in tensile strength and LOXL4] was observed only in breast cancer cells with a highly invasive/ structural integrity, and is essential for normal connective tissue metastatic phenotype but not in poorly invasive/nonmetastatic breast function, embryonic development, and wound healing (3, 5). Conse- cancer cells. LOX and LOXL2 showed the strongest association with quently, aberrant LOX expression or enzymatic activity leads to invasive potential in both highly invasive/metastatic breast cancer cell disease. Decreases in LOX expression or activity have been associated lines tested (MDA-MB-231 and Hs578T). To determine whether LOX with such heritable connective tissues disorders as Ehler-Danlos syn- is directly involved in breast cancer invasion, LOX antisense oligo- drome, cutis laxa, and Menkes’ syndrome (6–10). Increases in LOX nucleotides were transfected into MDA-MB-231 and Hs578T cells, and expression contribute to the development of fibrotic diseases such as found to inhibit invasion through a collagen IV/laminin/gelatin matrix in vitro compared with LOX sense oligonucleotide-treated and untreated arteriosclerosis, scleroderma, and liver cirrhosis, diseases that involve controls. In addition, treatment of MDA-MB-231 and Hs578T cells with connective tissue remodeling (11–14). ␤-aminopropionitrile (an irreversible inhibitor of LOX enzymatic Although the ECM maturation activity of LOX has long been activity) decreased invasive activity. Conversely, MCF-7 cells transfected with thought to be its sole function, more recent evidence implicates the the murine LOX gene demonstrated a 2-fold increase in invasiveness that was involvement of LOX in many important biological functions other reversible by the addition of ␤-aminopropionitrile in a dose-dependent than collagen/elastin cross-linking. LOX has been shown to induce manner. In addition, endogenous LOX mRNA expression was induced motility and migration in monocytes, vascular smooth muscle cells, when MCF-7 cells were cultured in the presence of fibroblast conditioned and fibroblasts (15–17). In addition, LOX may play a role in cell medium or conditioned matrix, suggesting a role for stromal fibroblasts in growth/differentiation as its expression is up-regulated by a number of LOX regulation in breast cancer cells. Moreover, the correlation of LOX growth factors and steroids (3, 4, 18). Moreover, a role for LOX in up-regulation and invasive/metastatic potential was additionally demon- strated in rat prostatic tumor cell lines, and human cutaneous and uveal transcriptional gene regulation and cellular transformation has also melanoma cell lines. These results provide substantial new evidence that been implicated as evidenced by localization of LOX protein and LOX is involved in cancer cell invasion. enzymatic activity to cell nuclei (19), potential utilization of histone H1 as a substrate (20), activation of the collagen III ␣1 promoter (21), and a putative tumor suppressor in nontumorigenic revertants of INTRODUCTION ras-transformed fibroblasts (reviewed in Ref. 3). In this study, we test the hypothesis that LOX expression contrib- In 2002, breast cancer is predicted to account for 31% of all of the utes to cellular invasive ability. The data demonstrate that LOX and diagnosed cancers in women, more than twice that of the second most other LOXL family members are specifically expressed by invasive/ common cancer (lung cancer; Ref. 1). Because metastasis is a major metastatic cancer cells compared with poorly invasive/nonmetastatic challenge in cancer management, it is critical to determine molecular cancer cells, and that modulation of LOX expression and function in markers that definitively distinguish tumors of nonmetastatic potential breast cancer cells affects in vitro cellular invasive activity. from those with metastatic potential. To identify such markers, a clearer understanding of the metastatic progression in breast cancer is MATERIALS AND METHODS required. To this end, we identified an up-regulation in LOX3 expres- sion in invasive/metastatic breast cancer cell lines compared with Cells and Culture Conditions. MCF-7 cells were kindly supplied by Dr. poorly invasive/nonmetastatic breast cancer cell lines (2). F. Miller (Karmanos Cancer Institute, Detroit, MI), and MDA-MB-231, T47D, and Hs578T cell lines were obtained from the American Type Culture Col- Received 2/15/02; accepted 5/23/02. lection (Manassas, VA). The cutaneous melanoma A375P and C8161 cell lines The costs of publication of this article were defrayed in part by the payment of page were kind gifts from Drs. I. J. Fidler (M.D. Anderson Cancer Center, Univer- charges. This article must therefore be hereby marked advertisement in accordance with sity of Texas, Houston, TX) and F. L. Meyskens, Jr. (University of California, 18 U.S.C. Section 1734 solely to indicate this fact. Irvine Cancer Center, Orange, CA), respectively. The uveal melanoma cell line 1 Supported by DAMD17-99-1-9225 (to D. A. K.), University of Iowa College of Medicine Grant (to D. A. K.), NIH/National Cancer Institute CA59702, Order of Eastern OCM-1 was kindly provided by Dr. J. Kan-Mitchell (Karmanos Cancer Insti- Star Breast Cancer Research Fund (to M. J. C. H.), NIH CA776580, and AR47713 tute), and M619 and C918 uveal melanoma cell lines were a kind gift from (to K. C.) Dr. K. Daniels (University of Iowa, Iowa City, IA). Human Rb foreskin 2 To whom requests for reprints should be addressed, at Department of Anatomy and fibroblasts were a generous gift from Dr. Gregory Goldberg (Washington Cell Biology, 1-100 BSB, University of Iowa, 51 Newton Road, Iowa City, IA 52242- Ј Ј Ј 1109. Phone: (319) 335-7655; Fax: (319) 335-7770; E-mail: dawn-kirschmann@ University, St. Louis, MO). Rat prostate cancer cell lines (5 P, 5 A, and 5 B) uiowa.edu. were derived and established from the R-3327 Dunning tumor in our labora- 3 The abbreviations used are: LOX, lysyl oxidase; Rb, foreskin fibroblast cell line; tory. Cell lines were maintained as described previously (22) and were har- ␤ ␤ LOXL, lysyl oxidase-like; ECM, extracellular matrix; APN, -aminopropionitrile; RT- vested when cultures were ϳ80% confluent. PCR, reverse transcriptase-PCR; MICS, membrane invasion culture system; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; hLOX, human LOX; mLOX, murine LOX; RNA Isolation. Total RNA was isolated from breast cancer cell lines using aa, amino acid; cm, conditioned medium; cmtx, conditioned matrix. TRIzol RNA isolation reagent (Invitrogen), according to manufacturer’s spec- 4478 Downloaded from cancerres.aacrjournals.org on September 26, 2021. © 2002 American Association for Cancer Research. LYSYL OXIDASE IN BREAST CANCER INVASION ifications. Polyadenylated RNA was isolated from total RNA by binding to centrifuged (2000 rpm, 10 min), filtered (0.22 ␮m) to remove residual cells and oligodeoxythymidine cellulose (Becton Dickinson, Bedford, MA), as de- cellular debris, and stored at Ϫ20°C until use. Conditioned matrices were scribed previously (23). generated by coating six-well tissue culture plates with 122 ␮g of rat tail Semiquantitative RT-PCR Analysis. Reverse transcription of total RNA collagen I (BD Bioscience) and polymerizing with 100% ethanol for 5 min at from breast, prostate, and melanoma cancer cell lines was performed using the room temperature. Matrices were rehydrated with 1 ml of PBS for 5 min at Advantage PCR kit according to manufacturer’s instructions (BD Clontech, room temperature, and 5 ϫ 105 Rb fibroblasts were added to each well in Palo Alto, CA). PCR amplification was performed with LOX- and LOXL2- complete medium and cultured for 3 days at 37°C, 5% CO2. Wells were treated Ј specific primers: hLOX [accession S78694 forward: 5 -GATCCTGCTGATC- with 20 mM NH4OH for 10 min at room temperature to remove the cells and CGCGACAA-3Ј (bases 500–520); reverse: 5Ј-GGGAGACCGTACTG- subsequently washed three times with sterile water and two times with PBS. GAAGTAGCCAGT-3Ј (bases 843–868)], rat LOX [accession U11038 MCF-7 cells (5 ϫ 105 cells/well) were then added to conditioned matrices for forward: 5Ј-CAACCCGGAAATTACATTCTAAAGGTCAGTG-3Ј (bases an additional 3 days. Total RNA was harvested by adding 1 ml of TRIzol 1376–1405); reverse: 5Ј-CCTTCCTCAGCACCAAGTGTATCC-3Ј (bases reagent to each well. 1986–2009)], and hLOXL2 [accession NM_002318 forward: 5Ј-GGCCGC- Ј Ј CACGCGTG GATC-3 (bases 2093–2110); reverse: 5 -CCCAAGGGTCAG- RESULTS GTAGCAGCCCC-3Ј (bases 2752–2774)]. Annealing temperature and number of amplification cycles were optimized at 64°C and 30 cycles for LOX, and LOX Expression in Breast Cancer Cell Lines.
Load more

Recommended publications
  • Lysyl Oxidase Family Members in Urological Tumorigenesis and Fibrosis
    www.oncotarget.com Oncotarget, 2018, Vol. 9, (No. 28), pp: 20156-20164 Review Lysyl oxidase family members in urological tumorigenesis and fibrosis Tao Li1,2, Changjing Wu1, Liang Gao3, Feng Qin1, Qiang Wei2 and Jiuhong Yuan1,2 1The Andrology Laboratory, West China Hospital, Sichuan University, Chengdu, Sichuan, China 2Department of Urology, West China Hospital, Sichuan University, Chengdu, Sichuan, China 3Department of Urology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, China Correspondence to: Jiuhong Yuan, email: [email protected] Keywords: lysyl oxidase; tumorigenesis; fibrosis; urological cancer; collagen Received: September 29, 2017 Accepted: March 11, 2018 Published: April 13, 2018 Copyright: Li et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License 3.0 (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT Lysyl oxidase (LOX) is an extracellular copper-dependent monoamine oxidase that catalyzes crosslinking of soluble collagen and elastin into insoluble, mature fibers. Lysyl oxidase-like proteins (LOXL), LOX isozymes with partial structural homology, exhibit similar catalytic activities. This review summarizes recent findings describing the roles of LOX family members in urological cancers and fibrosis. LOX/ LOXL play key roles in extracellular matrix stability and integrity, which is essential for normal female pelvic floor function. LOX/LOXL inhibition may reverse kidney fibrosis and ischemic priapism. LOX and LOXL2 reportedly promote kidney carcinoma tumorigenesis, while LOX, LOXL1 and LOXL4 suppress bladder cancer growth. Multiple studies agree that the LOX propeptide may suppress tumor growth, but the role of LOX in prostate cancer remains controversial.
    [Show full text]
  • The Function and Mechanisms of Action of LOXL2 in Cancer (Review)
    1200 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 36: 1200-1204, 2015 The function and mechanisms of action of LOXL2 in cancer (Review) LINGHONG WU and YING ZHU Department of Infectious Diseases, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China Received May 31, 2015; Accepted August 26, 2015 DOI: 10.3892/ijmm.2015.2337 Abstract. The lysyl oxidase (LOX) family is comprised of five copper‑dependent amine oxidase, and its main role is to members, and some members have recently emerged as impor- catalyze the covalent cross‑link of collagen and elastin in tant regulators of tumor progression. Among these, at present, the extracellular matrix (ECM). This occurs through the LOX‑like (LOXL)2 is the prototypical LOX and the most oxidative deamination of peptidyl lysine residues in ECM comprehensively studied member. A growing body of evidence components (1,2). Each member of the LOX family has a has implicated LOXL2 in the promotion of cancer cell inva- highly conserved carboxyl (C)‑terminal domain that contains a sion, metastasis and angiogenesis, as well as in the malignant copper‑binding motif, lysine tyrosylquinone (LTQ) residues and transformation of solid tumors. Moreover, a high expression of a cytokine receptor‑like (CRL) domain, which is essential for LOXL2 is associated with a poor prognosis. These data have catalytic activity (3). The amino‑terminal regions are different piqued the interest of a number of researchers and research and are thought to be important in protein‑protein interactions. groups, who have identified LOXL2 as a strong target candidate The prodomains in LOX and LOXL1 enable their secretion in the development of inhibitors for use as functional and effi- as inactive proenzymes, which are then activated extracel- cacious tumor therapeutics.
    [Show full text]
  • Genetic Analyses of Human Fetal Retinal Pigment Epithelium Gene Expression Suggest Ocular Disease Mechanisms
    ARTICLE https://doi.org/10.1038/s42003-019-0430-6 OPEN Genetic analyses of human fetal retinal pigment epithelium gene expression suggest ocular disease mechanisms Boxiang Liu 1,6, Melissa A. Calton2,6, Nathan S. Abell2, Gillie Benchorin2, Michael J. Gloudemans 3, 1234567890():,; Ming Chen2, Jane Hu4, Xin Li 5, Brunilda Balliu5, Dean Bok4, Stephen B. Montgomery 2,5 & Douglas Vollrath2 The retinal pigment epithelium (RPE) serves vital roles in ocular development and retinal homeostasis but has limited representation in large-scale functional genomics datasets. Understanding how common human genetic variants affect RPE gene expression could elu- cidate the sources of phenotypic variability in selected monogenic ocular diseases and pin- point causal genes at genome-wide association study (GWAS) loci. We interrogated the genetics of gene expression of cultured human fetal RPE (fRPE) cells under two metabolic conditions and discovered hundreds of shared or condition-specific expression or splice quantitative trait loci (e/sQTLs). Co-localizations of fRPE e/sQTLs with age-related macular degeneration (AMD) and myopia GWAS data suggest new candidate genes, and mechan- isms by which a common RDH5 allele contributes to both increased AMD risk and decreased myopia risk. Our study highlights the unique transcriptomic characteristics of fRPE and provides a resource to connect e/sQTLs in a critical ocular cell type to monogenic and complex eye disorders. 1 Department of Biology, Stanford University, Stanford, CA 94305, USA. 2 Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA. 3 Program in Biomedical Informatics, Stanford University School of Medicine, Stanford 94305 CA, USA. 4 Department of Ophthalmology, Jules Stein Eye Institute, UCLA, Los Angeles 90095 CA, USA.
    [Show full text]
  • LOXL1 Confers Antiapoptosis and Promotes Gliomagenesis Through Stabilizing BAG2
    Cell Death & Differentiation (2020) 27:3021–3036 https://doi.org/10.1038/s41418-020-0558-4 ARTICLE LOXL1 confers antiapoptosis and promotes gliomagenesis through stabilizing BAG2 1,2 3 4 3 4 3 1 1 Hua Yu ● Jun Ding ● Hongwen Zhu ● Yao Jing ● Hu Zhou ● Hengli Tian ● Ke Tang ● Gang Wang ● Xiongjun Wang1,2 Received: 10 January 2020 / Revised: 30 April 2020 / Accepted: 5 May 2020 / Published online: 18 May 2020 © The Author(s) 2020. This article is published with open access Abstract The lysyl oxidase (LOX) family is closely related to the progression of glioma. To ensure the clinical significance of LOX family in glioma, The Cancer Genome Atlas (TCGA) database was mined and the analysis indicated that higher LOXL1 expression was correlated with more malignant glioma progression. The functions of LOXL1 in promoting glioma cell survival and inhibiting apoptosis were studied by gain- and loss-of-function experiments in cells and animals. LOXL1 was found to exhibit antiapoptotic activity by interacting with multiple antiapoptosis modulators, especially BAG family molecular chaperone regulator 2 (BAG2). LOXL1-D515 interacted with BAG2-K186 through a hydrogen bond, and its lysyl 1234567890();,: 1234567890();,: oxidase activity prevented BAG2 degradation by competing with K186 ubiquitylation. Then, we discovered that LOXL1 expression was specifically upregulated through the VEGFR-Src-CEBPA axis. Clinically, the patients with higher LOXL1 levels in their blood had much more abundant BAG2 protein levels in glioma tissues. Conclusively, LOXL1 functions as an important mediator that increases the antiapoptotic capacity of tumor cells, and approaches targeting LOXL1 represent a potential strategy for treating glioma.
    [Show full text]
  • Identification of the Involvement of LOXL4 in Generation of Keratocystic Odontogenic Tumors by RNA-Seq Analysis
    International Journal of Oral Science (2013) 6, 31–38 ß 2013 WCSS. All rights reserved 1674-2818/13 www.nature.com/ijos ORIGINAL ARTICLE Identification of the involvement of LOXL4 in generation of keratocystic odontogenic tumors by RNA-Seq analysis Wei-Peng Jiang1, Zi-Han Sima1, Hai-Cheng Wang1, Jian-Yun Zhang1, Li-Sha Sun2, Feng Chen2 and Tie-Jun Li1 Keratocystic odontogenic tumors (KCOT) are benign, locally aggressive intraosseous tumors of odontogenic origin. KCOT have a higher stromal microvessel density (MVD) than dentigerous cysts (DC) and normal oral mucosa. To identify genes in the stroma of KCOT involved in tumor development and progression, RNA sequencing (RNA-Seq) was performed using samples from KCOT and primary stromal fibroblasts isolated from gingival tissues. Seven candidate genes that possess a function potentially related to KCOT progression were selected and their expression levels were confirmed by quantitative PCR, immunohistochemistry and enzyme-linked immunosorbent assay. Expression of lysyl oxidase-like 4 (LOXL4), the only candidate gene that encodes a secreted protein, was enhanced at both the mRNA and protein levels in KCOT stromal tissues and primary KCOT stromal fibroblasts compared to control tissues and primary fibroblasts (P,0.05). In vitro, high expression of LOXL4 could enhance proliferation and migration of the human umbilical vein endothelial cells (HUVECs). There was a significant, positive correlation between LOXL4 protein expression and MVD in stroma of KCOT and control tissues (r50.882). These data suggest that abnormal expression of LOXL4 of KCOT may enhance angiogenesis in KCOT, which may help to promote the locally aggressive biological behavior of KCOT.
    [Show full text]
  • Cell-Deposited Matrix Improves Retinal Pigment Epithelium Survival on Aged Submacular Human Bruch’S Membrane
    Retinal Cell Biology Cell-Deposited Matrix Improves Retinal Pigment Epithelium Survival on Aged Submacular Human Bruch’s Membrane Ilene K. Sugino,1 Vamsi K. Gullapalli,1 Qian Sun,1 Jianqiu Wang,1 Celia F. Nunes,1 Noounanong Cheewatrakoolpong,1 Adam C. Johnson,1 Benjamin C. Degner,1 Jianyuan Hua,1 Tong Liu,2 Wei Chen,2 Hong Li,2 and Marco A. Zarbin1 PURPOSE. To determine whether resurfacing submacular human most, as cell survival is the worst on submacular Bruch’s Bruch’s membrane with a cell-deposited extracellular matrix membrane in these eyes. (Invest Ophthalmol Vis Sci. 2011;52: (ECM) improves retinal pigment epithelial (RPE) survival. 1345–1358) DOI:10.1167/iovs.10-6112 METHODS. Bovine corneal endothelial (BCE) cells were seeded onto the inner collagenous layer of submacular Bruch’s mem- brane explants of human donor eyes to allow ECM deposition. here is no fully effective therapy for the late complications of age-related macular degeneration (AMD), the leading Control explants from fellow eyes were cultured in medium T cause of blindness in the United States. The prevalence of only. The deposited ECM was exposed by removing BCE. Fetal AMD-associated choroidal new vessels (CNVs) and/or geo- RPE cells were then cultured on these explants for 1, 14, or 21 graphic atrophy (GA) in the U.S. population 40 years and older days. The explants were analyzed quantitatively by light micros- is estimated to be 1.47%, with 1.75 million citizens having copy and scanning electron microscopy. Surviving RPE cells from advanced AMD, approximately 100,000 of whom are African explants cultured for 21 days were harvested to compare bestro- American.1 The prevalence of AMD increases dramatically with phin and RPE65 mRNA expression.
    [Show full text]
  • Transdifferentiation of Human Mesenchymal Stem Cells
    Transdifferentiation of Human Mesenchymal Stem Cells Dissertation zur Erlangung des naturwissenschaftlichen Doktorgrades der Julius-Maximilians-Universität Würzburg vorgelegt von Tatjana Schilling aus San Miguel de Tucuman, Argentinien Würzburg, 2007 Eingereicht am: Mitglieder der Promotionskommission: Vorsitzender: Prof. Dr. Martin J. Müller Gutachter: PD Dr. Norbert Schütze Gutachter: Prof. Dr. Georg Krohne Tag des Promotionskolloquiums: Doktorurkunde ausgehändigt am: Hiermit erkläre ich ehrenwörtlich, dass ich die vorliegende Dissertation selbstständig angefertigt und keine anderen als die von mir angegebenen Hilfsmittel und Quellen verwendet habe. Des Weiteren erkläre ich, dass diese Arbeit weder in gleicher noch in ähnlicher Form in einem Prüfungsverfahren vorgelegen hat und ich noch keinen Promotionsversuch unternommen habe. Gerbrunn, 4. Mai 2007 Tatjana Schilling Table of contents i Table of contents 1 Summary ........................................................................................................................ 1 1.1 Summary.................................................................................................................... 1 1.2 Zusammenfassung..................................................................................................... 2 2 Introduction.................................................................................................................... 4 2.1 Osteoporosis and the fatty degeneration of the bone marrow..................................... 4 2.2 Adipose and bone
    [Show full text]
  • Reduced Lysyl Oxidase-Like (LOXL) in the Skin and Heart
    See related Commentary on page vi Progressive Hair Loss and Myocardial Degeneration in Rough Coat Mice: Reduced Lysyl Oxidase-Like (LOXL) in the Skin and Heart Kimiko Hayashi,à Tongyu Cao,à Howard Passmore,w Claude Jourdan-Le Saux,à Ben Fogelgren,à Subarna Khan,w Ian Hornstra,z Youngho Kim,y Masando Hayashi,à and Katalin Csiszarà ÃJohn A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, Hawaii, USA; wDepartment of Genetics, Rutgers University, Piscataway, New Jersey, USA; zDepartment of Medicine, Division of Dermatology, Barnes-Jewish Hospital, Washington University School of Medicine, St Louis, Missouri, USA; yGenome Research Center for Birth Defects and Genetic Diseases, Asan Medical Center, Seoul, Korea The rough coat (rc) is a spontaneous recessive mutation in mice. To identify the mutated gene, we have char- acterized the rc phenotype and initiated linkage mapping. The rc mice show growth retardation, cyclic and pro- gressive hair loss, hyperplastic epidermis, abnormal hair follicles, cardiac muscle degeneration, and reduced amount of collagen and elastin in the skin and heart. The rc locus was mapped at 32.0 cM on chromosome 9, close to the loxl gene. Lysyl oxidase-like (LOXL) protein is a novel copper-containing amine oxidase that is required for the cross-linking of elastin and collagen in vitro. LOXL is expressed at high levels in the skin and heart, where the rc mice show strong phenotype. The expression pattern and the genetic proximity to rc suggested loxl as a potential candidate gene. In rc mice, the loxl mRNA was reduced in the skin and the LOXL protein in the heart, dermis, atrophic hair follicles, and sebaceous glands.
    [Show full text]
  • LAMA2 and LOXL4 Are Candidate FSGS Genes
    LAMA2 and LOXL4 Are Candidate FSGS Genes Saidah Hack Toronto General Hospital Poornima Vijayan University of Toronto Tony Yao Toronto General Hospital Mohammad Azfar Qureshi Toronto General Hospital Andrew Paterson SickKids: The Hospital for Sick Children Rohan John Toronto General Hospital Bernard Davenport Wellcome Centre Cell-Matrix Research: Wellcome Trust Centre for Cell Matrix Research Rachel Lennon Wellcome Centre Cell-Matrix Research: Wellcome Trust Centre for Cell Matrix Research York Pei Toronto General Hospital Moumita Barua ( [email protected] ) Toronto General Hospital https://orcid.org/0000-0003-0628-9071 Research article Keywords: Hereditary FSGS, basement membrane, LAMA2, LOXL4 Posted Date: February 26th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-257952/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/8 Abstract Background: Focal and segmental glomerulosclerosis (FSGS) is a histologic pattern of injury that characterizes a wide spectrum of diseases. Many genetic causes have been identied in FSGS but even in families with comprehensive testing, a signicant proportion remain unexplained. Results: In one FSGS family with 6 affected relatives, we performed linkage analysis and whole exome sequencing (WES). Linkage analysis narrowed the disease gene(s) to within 3% of the genome. Whole exome sequencing revealed 5 heterozygous rare variants, which were sequenced in 11 relatives where DNA was available. Two of these variants, in LAMA2 and LOXL4, remained as candidates after segregation analysis and encode extracellular matrix proteins of the glomerulus. Renal biopsies showed classic segmental sclerosis/hyalinosis lesion on a background of mild mesangial hypercellularity.
    [Show full text]
  • LOXL3-Sv2, a Novel Variant of Human Lysyl Oxidase-Like 3 (LOXL3), Functions As an Amine Oxidase
    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 39: 719-724, 2017 LOXL3-sv2, a novel variant of human lysyl oxidase-like 3 (LOXL3), functions as an amine oxidase CHANKYU JEONG and YOUNGHO KIM Department of Biochemistry, Wonkwang University School of Medicine, Institute of Wonkwang Medical Science, Iksan, Jeonbuk 570-749, Republic of Korea Received May 23, 2016; Accepted January 13, 2017 DOI: 10.3892/ijmm.2017.2862 Abstract. Human lysyl oxidase-like 3 (LOXL3) functions paralogues (LOX, LOXL, LOXL2, LOXL3 and LOXL4) as a copper-dependent amine oxidase toward collagen and contain a copper-binding domain, residues for lysyl-tyrosyl elastin. The LOXL3 protein contains four scavenger receptor quinone (LTQ) and a cytokine receptor-like (CRL) domain cysteine-rich (SRCR) domains in the N-terminus in addi- in the C-terminus. In addition, four repeated copies of scav- tion to the C-terminal characteristic domains of the lysyl enger receptor cysteine-rich (SRCR) domains are found in the oxidase (LOX) family, such as a copper-binding domain, a N-terminus of only LOXL2, LOXL3 and LOXL4, suggesting cytokine receptor-like domain and residues for the lysyl-tyrosyl novel functional roles assigned to these three members (4-6). quinone cofactor. Using BLASTN searches, we identified a LOXL3 has been reported to be associated with a diverse novel variant of LOXL3 (termed LOXL3-sv2), which lacked range of diseases in both humans and animals. LOXL3 was the sequences corresponding to exons 4 and 5 of LOXL3. The reported to interact with Snail to downregulate the expression LOXL3-sv2 mRNA is at least 2,398 bp in length, encoding a of E-cadherin in carcinoma progression (7), to induce the 608 amino acid-long polypeptide with a calculated molecular autophagy of chondrocytes in osteoarthritis (8) and to play an mass of 67.4 kDa.
    [Show full text]
  • Hypoxia-Inducible Factor 1 Is a Master Regulator of Breast Cancer Metastatic Niche Formation
    Hypoxia-inducible factor 1 is a master regulator of breast cancer metastatic niche formation Carmen Chak-Lui Wonga,b, Daniele M. Gilkesa,b, Huafeng Zhanga,c,d, Jasper Chena,b, Hong Weia,b, Pallavi Chaturvedia,b, Stephanie I. Fraleye, Chun-Ming Wongf,g, Ui-Soon Khoog, Irene Oi-Lin Ngf,g, Denis Wirtze, and Gregg L. Semenzaa,b,c,1 aVascular Program, Institute for Cell Engineering, bMcKusick–Nathans Institute of Genetic Medicine, and cDepartment of Oncology, The Johns Hopkins University School of Medicine, Baltimore, MD 21205; eDepartment of Chemical and Biomolecular Engineering, The Johns Hopkins University, Baltimore, MD 21218; fState Key Laboratory for Liver Research and gDepartment of Pathology, Li Ka shing Faculty of Medicine, University of Hong Kong, Hong Kong; and dSchool of Life Science, University of Science and Technology of China, Hefei, Anhui, China Contributed by Gregg L. Semenza, August 17, 2011 (sent for review July 25, 2011) Primary tumors facilitate metastasis by directing bone marrow- (vol/vol/vol; 1% O2). Each cell line exhibited a different pattern derived cells (BMDCs) to colonize the lungs before the arrival of of expression in response to hypoxia (Fig. 1A and Fig. S1 A–C). cancer cells. Here, we demonstrate that hypoxia-inducible factor 1 In MDA-231, LOX and LOXL4 were induced by hypoxia, (HIF-1) is a critical regulator of breast cancer metastatic niche whereas in MDA-435, LOXL2 was induced by hypoxia. LOX was formation through induction of multiple members of the lysyl hypoxia-induced in MDA-231, as previously reported (8), but not B oxidase (LOX) family, including LOX, LOX-like 2, and LOX-like 4, in MDA-435 (Fig.
    [Show full text]
  • Lysyl Oxidase Modulates the Osteoblast Differentiation of Primary Mouse Calvaria Cells
    1664 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 36: 1664-1670, 2015 Lysyl oxidase modulates the osteoblast differentiation of primary mouse calvaria cells ANJALI SHARMA-BHANDARI1, SUN-HYANG PARK1, JU-YOUNG KIM2, JAEMIN OH2 and YOUNGHO KIM1 Departments of 1Biochemistry and 2Anatomy, School of Medicine, Institute of Wonkwang Medical Science, Wonkwang University, Iksan, Jeollabuk-do 570-749, Republic of Korea Received April 23, 2015; Accepted October 14, 2015 DOI: 10.3892/ijmm.2015.2384 Abstract. Lysyl oxidase (LOX) is an extracellular amine matrix functions as a three-dimensional scaffold for organizing oxidase that mediates the formation of collagen fibers. Thus mineral deposition in bone (1). Many studies have reported that far, five LOX family genes [LOX, lysyl oxidase-like (LOXL)1, altered cross-linking of collagen type I exerts significant effects LOXL2, LOXL3 and LOXL4] have been identified in humans, on the differentiation and mineralization of various cell types, each encoding the characteristic C-terminal domains that are including endometrial cells, smooth muscle cells and mammary required for amine oxidase activity. During osteoblastogenesis, cells (2-4). The aberrant cross-linking of collagen fibrils has collagen fibers function as a three-dimensional scaffold for orga- been observed in a number of bone disorders, including osteo- nizing mineral deposition. In this study, to assess the functional porosis, osteopetrosis and diabetes-related bone disease (5-7). roles of the LOX family members in osteoblastogenesis, we Lysyl oxidase (LOX) is a secreted, copper-dependent amine investigated the temporal expression of these genes as a function oxidase that plays a key role in maintaining the integrity of of phenotypic development during the osteoblast differentiation connective tissue by modulating the cross-linking of collagen of primary cultured mouse calvaria cells.
    [Show full text]