DOCSLIB.ORG

Lncrna DNAJC3-AS1 Functions As Oncogene in Renal Cell Carcinoma Via Regulation of the Mir-27A-3P/PRDM14 Axis

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Lncrna DNAJC3-AS1 Functions As Oncogene in Renal Cell Carcinoma Via Regulation of the Mir-27A-3P/PRDM14 Axis European Review for Medical and Pharmacological Sciences 2021; 25: 1291-1301 LncRNA DNAJC3-AS1 functions as oncogene in renal cell carcinoma via regulation of the miR-27a-3p/PRDM14 axis X.-Y. NA, X.-Q. HU, Y. ZHAO, C.-H. HU, X.-S. SHANG Department of Neurology, People’s Hospital of Zhenhai District, Ningbo, China (testis-associated highly conserved oncogenic long Abstract. – OBJECTIVE: Renal cell carcino- ma (RCC) is one of the most common urologi- non-coding RNA); DNAJC3-AS1 (DNAJ/HSP40 homo- cal malignancies worldwide. Although great ad- log, subfamily C, member 3, anti-sense 1); RPDM14 vances have been made in the diagnosis and (PR domain-containing membrane 14); SNHG12 (small management of RCC, its prognosis remains un- nucleolar RNA host gene 12); CCK-8 (Cell Count Kit- satisfactory. Long noncoding RNAs (lncRNAs) 8) ZEB1-AS1 (zinc-finger E-box binding homeobox 1, have been found to be essential factors in the anti-sense 1); FBS (Fetal Bovine Serum); DMEM (Dul- initiation and development of cancer. The cur- becco’s Modified Eagle’s Medium); DEVD-pNA (Asp- rent study aimed to measure the expression and Glu-Val-Asp-chromophore p-nitroanilide); ELISA (en- functions of lncRNA DNAJC3-AS1 in the pro- zyme-linked immunosorbent assay); SDS-PAGE (sodi- gression of clear cell RCC (ccRCC). um dodecyl sulfate-polyacrylamide gel electrophoresis); PATIENTS AND METHODS: The expression RIPA buffer (Radio-Immunoprecipitation assay buffer); of lncRNA DNAJC3-AS1 was detected in 30 PVDF (Polyvinylidene fluoride). pairs of ccRCC tissues and in cell lines by RT- PCR, and its prognostic association with ccRCC was evaluated by the Kaplan-Meier method. The proliferation, migration, invasion and apopto- Introduction sis of ccRCC cells were measured after silenc- ing DNAJC3-AS1. The interaction between DNA- Renal cell carcinoma (RCC) is a common kid- JC3-AS1, miR-27a-3p and PRDM14 was identi- ney carcinoma that accounts for 80-90% of adult fied by Dual-Luciferase reporter assay. The pro- 1 tein levels were measured by Western blotting. renal malignant tumors . Worldwide, approxima- RESULTS: The expression of DNAJC3-AS1 tely 270,000 new cases of RCC are diagnosed and was upregulated in ccRCC tissues and cell lines over ten thousand RCC patients die each year2. compared to their normal counterparts. In vi- Clear cell RCC (ccRCC) accounts for approxi- tro, silencing DNAJC3-AS1 reduced the prolif- mately 90% of all RCCs in terms of histological eration, migration and invasion of ccRCC cells. subtype; therefore, it is one of the most lethal Downregulation of DNAJC3-AS1 also led to the apoptosis of ccRCC cells. Moreover, we also urological malignancies. Currently, the diagnosis found that DNAJC3-AS1 acted as a sponge of of ccRCC primarily depends on computed tomo- miR-27a-3p and identified PRDM14 as a target of graphy scans and magnetic resonance imaging. miR-27a-3p. Because RCC is insensitive to radiotherapy or CONCLUSIONS: LncRNA DNAJC3-AS1 acts chemotherapy, the treatment of early RCC mainly as an oncogene and plays an essential role in relies on surgery. Although the treatment of RCC the tumorigenesis of ccRCC, possibly via the has greatly improved in the last two decades, the regulation of the miR-27a-3p/PRDM14 axis. prognosis of RCC remains poor. Moreover, there Key Words: is still a lack of effective biomarkers for predi- Renal cell carcinoma, LncRNA DNAJC3-AS1, MiR- cting clinical progression and outcome in ccRCC 27a-3p, PRDM14. patients. Thus, it is urgent to identify effective biomarkers and unveil the molecular mechanisms of ccRCC progression. Abbreviations In recent years, long noncoding RNA (lncR- RCC (renal cell carcinoma); ccRCC (clear cell renal NA), which was initially considered the “junk” of cell carcinoma); RT-PCR (real-time reverse transcrip- the genome, has been found to play an essential tion PCR); lncRNA (long non coding RNA); THOR role in various biological activities3. Some stu- Corresponding Author: Shang Xiaosong, MD; e-mail: [email protected] 1291 X.-Y. Na, X.-Q. Hu, Y. Zhao, C.-H. Hu, X.-S. Shang dies4-6 have shown that lncRNAs are associated obtained from all patients. The Ethics Commit- with the development of various cancers, such as tee of the hospital reviewed and approved this lung cancer, breast cancer, colorectal cancer and study. gastric cancer. It has been reported that various lncRNAs are involved in the tumorigenesis of Cell Culture and Transfection RCC. For instance, lncRNA THOR is upregu- ccRCC cell lines (769-P, ACHN, Caki-1, 786- lated in RCC tissues and cell lines. Silencing of O) and a normal renal proximal tubular epithelial THOR inhibited the growth, migration and inva- cell line (HK-2) were purchased from Shanghai sion of RCC cells7. LncRNA SNHG12 was also Bank of Cells (Shanghai, China). Cells were cul- overexpressed in ccRCC tissues and cell lines. tured in DMEM (HyClone, South-Logan, UT, Inhibition of SNHG12 repressed the viability and USA) supplemented with 10% FBS (fetal bovine mobility of ccRCC cells8. serum, penicillin; 100 U/ml) and streptomycin Notably, antisense partners of protein-coding (100 μg/mL; Gibco, Carlsbad, CA, USA). All cel- genes are one type of lncRNA; these include ls were cultured in a humidified atmosphere of 9-11 DLX6-AS1, ZEB1-AS1 and TP73-AS1 . LncR- 5% CO2 at 37°C. The siRNAs for DNAJC3-AS1 NA DNAJC3-AS1 is a relatively novel lncRNA (si-DNAJC3-AS1 #1, si-DNAJC3-AS1 #2, si-D- that is located on chromosome 13q32.1 and has NAJC3-AS1 #3) and PRDM14 (si-PRDM14), and 2 exons. DNAJC3, also known as P58, belongs miR-27a-3p mimics, miR-27a-3p inhibitors, con- to the heat shock protein family, which is a very trol siRNA (si-NC), and miR-NC mimics were conserved protein that is widely expressed in hu- synthesized by GenePharma Ltd (Suzhou, Chi- man and animal cells12. It is well documented that na). The coding region of PRDM14 was ampli- DNAJC3 can provide protective effects against fied from cDNA and cloned into pcDNA3.1 (pc- various kinds of damage in cells12,13. Howe- DNA3.1 PRDM14). Transfection was performed ver, there is little investigation about the role of using Lipofectamine 3000 (Life Technologies, DNAJC3-AS1 in ccRCC. Carlsbad, CA, USA) according to the manufactu- In the present study, the correlation between rer’s instructions. DNAJC3-AS1 and clinical data in RCC patients was investigated. Furthermore, we studied the in Cell Proliferation Assay vitro effects of DNAJC3-AS1 on ccRCC cells and The proliferation of cells was measured by the elucidated its underlying mechanisms. Cell Counting Kit-8 (CCK-8; Solarbio, Beijing, China) assay according to the manufacturer’s in- structions. Briefly, cells were plated in 96-well Patients and Methods culture plates at a density of 1 ×104 cells/well and cultured in fresh medium mixed with CCK-8 at a Patients and Tissues Collection ratio of 10:1 for 2 h. The cell density was measu- In total, 30 pairs of ccRCC and adjacent nor- red with a microplate reader (Biotek, Winooski, mal tissues were collected during nephrectomy VT, USA) at 490 nm. of ccRCC patients who did not receive any preo- perative treatment at the hospital. The inclusion RNA Purification and RT-PCR criteria were as follows: no history of other ma- RNA was extracted from cells or tissues using lignant tumors, no history of anticancer therapy, TRIzol (Life Technologies, Carlsbad, CA, USA). pathologically proven ccRCC and radical or par- The concentration and quality of RNA were as- tial nephrectomy as the treatment. The exclusion sayed using a NanoDrop 2000 Spectrophotometer criteria were as follows: mixed histological type (Thermo Scientific, Waltham, MA, USA). One of primary renal cancer, larger area of necro- microgram of total RNA was reverse transcribed sis and hemorrhage influencing the collection into cDNA using the High Capacity cDNA Rever- of representative area in samples, and death of se Transcriptase Kit (Life Technologies, Carlsbad, the patient within the first month after surgery. CA, USA). qRT-PCR was carried out using SYBR The diagnosis was histologically and pathologi- GreenTM qPCR SuperMix Universal (Life Techno- cally confirmed by 2 experienced pathologists. logies, Carlsbad, CA, USA) on an Applied Biosy- All tumors were staged according to the AJCC stems 7500 Detection System (Applied Biosystems, 7th system14. All samples were instantly frozen Foster City, CA, USA) to measure the expres- in liquid nitrogen after resection and then main- sion of DNAJC3-AS1, miR-27a-3p, PRDM14, U6 tained in storage. Written informed consent was and GAPDH. The following primers were used: 1292 LncRNA DNAJC3-AS1 functions as oncogene in RCC via regulation of the miR-27a-3p/PRDM14 axis DNAJC3-AS1 forward: 5′-AGCGATTGTG- for caspase-3 activity was performed in a 96-well GAAGACCCTG-3′ and reverse: 5′-ATTTCCC- plate. After transfection for 24 h, cells were col- CTGGTAAGCGCAA-3′; miR-27a-3p forward: lected and lysed. Fifty microliters of lysates were 5′-TTCACAGTGGCTAAGTTCCGC-3′ and 5′- added into each well. A 50 μl aliquot of 2× re- AGGG CTTAGCTGCTTGTGAGCA-3′; PRDM14 action buffer was added to each sample. Then, forward: 5’-TGAGCCTTCAGGTCACAGAG-3’ 200 μM DEVD-pNA was added as the caspase-3 and reverse 5’-ATTTCCTATCGCCCTTGTCC-3’; substrate. The reaction mixtures were mixed and U6 forward: 5′-CTCGCTTCGGCAGCACA-3′ and incubated at 37°C for 2 h. Then, the absorbance reverse 5′-AACGCTTCACGAATTTGCGT-3′; was measured by a microplate reader (Biotek, and GAPDH forward: 5′-GGTGAAGGTCGGA- Winooski, VT, USA) at 405 nm. GTCAACG-3′ and reverse: 5′-CAAAGTTGTCA- TGGATGTACC-3′. The PCR conditions were as Apoptosis Assay follows: 95°C for 10 min, 40 cycles of 95°C for 15 The apoptosis of cells was performed with s, and 60°C for 60 s.
Load more

Recommended publications
  • Deletion of Endoplasmic Reticulum Stress-Responsive Co-Chaperone P58ipk Protects Mice from Diet-Induced Steatohepatitis
    Zurich Open Repository and Archive University of Zurich Main Library Strickhofstrasse 39 CH-8057 Zurich www.zora.uzh.ch Year: 2018 Deletion of endoplasmic reticulum stress-responsive co-chaperone p58IPK protects mice from diet-induced steatohepatitis Bandla, Harikrishna ; Dasgupta, Debanjali ; Mauer, Amy S ; Nozickova, Barbora ; Kumar, Swarup ; Hirsova, Petra ; Graham, Rondell P ; Malhi, Harmeet DOI: https://doi.org/10.1111/hepr.13052 Posted at the Zurich Open Repository and Archive, University of Zurich ZORA URL: https://doi.org/10.5167/uzh-144890 Journal Article Accepted Version Originally published at: Bandla, Harikrishna; Dasgupta, Debanjali; Mauer, Amy S; Nozickova, Barbora; Kumar, Swarup; Hirsova, Petra; Graham, Rondell P; Malhi, Harmeet (2018). Deletion of endoplasmic reticulum stress-responsive co-chaperone p58IPK protects mice from diet-induced steatohepatitis. Hepatology Research, 48(6):479- 494. DOI: https://doi.org/10.1111/hepr.13052 Deletion of endoplasmic reticulum stress-responsive co-chaperone p58IPK protects mice from diet-induced steatohepatitis Harikrishna Bandla1, Debanjali Dasgupta1, Amy S. Mauer1, Barbora Nozickova2, Swarup Kumar3, Petra Hirsova1, Rondell P. Graham4, Harmeet Malhi1* 1. Division of Gastroenterology and Hepatology, Mayo Clinic, Rochester, MN 2. Universitatsspital Zurich, 8096, Ramistrasse 100, Zurich, Switzerland 3. Department of Medicine, Saint Vincent Hospital, 123 Summer St, Worcester, MA 4. Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN Corresponding author: Harmeet Malhi, M.B.B.S. Associate Professor of Medicine and Physiology Mayo Clinic College of Medicine 200 First Street SW Rochester, MN 55905 Tel: 507 284 0686 Fax: 507 284 0762 Email: [email protected] Funding: This work was supported by DK 97178, DK107402 and DK111378 (H.M.), the Robert and Elizabeth Strickland Career Development Award from the Division of Endocrinology (H.M.), the Gilead Sciences Research Scholars Program in Liver Disease (H.M.) and the Palumbo Foundation (H.M.), the Edward C.
    [Show full text]
  • Computational Genome-Wide Identification of Heat Shock Protein Genes in the Bovine Genome [Version 1; Peer Review: 2 Approved, 1 Approved with Reservations]
    F1000Research 2018, 7:1504 Last updated: 08 AUG 2021 RESEARCH ARTICLE Computational genome-wide identification of heat shock protein genes in the bovine genome [version 1; peer review: 2 approved, 1 approved with reservations] Oyeyemi O. Ajayi1,2, Sunday O. Peters3, Marcos De Donato2,4, Sunday O. Sowande5, Fidalis D.N. Mujibi6, Olanrewaju B. Morenikeji2,7, Bolaji N. Thomas 8, Matthew A. Adeleke 9, Ikhide G. Imumorin2,10,11 1Department of Animal Breeding and Genetics, Federal University of Agriculture, Abeokuta, Nigeria 2International Programs, College of Agriculture and Life Sciences, Cornell University, Ithaca, NY, 14853, USA 3Department of Animal Science, Berry College, Mount Berry, GA, 30149, USA 4Departamento Regional de Bioingenierias, Tecnologico de Monterrey, Escuela de Ingenieria y Ciencias, Queretaro, Mexico 5Department of Animal Production and Health, Federal University of Agriculture, Abeokuta, Nigeria 6Usomi Limited, Nairobi, Kenya 7Department of Animal Production and Health, Federal University of Technology, Akure, Nigeria 8Department of Biomedical Sciences, Rochester Institute of Technology, Rochester, NY, 14623, USA 9School of Life Sciences, University of KwaZulu-Natal, Durban, 4000, South Africa 10School of Biological Sciences, Georgia Institute of Technology, Atlanta, GA, 30032, USA 11African Institute of Bioscience Research and Training, Ibadan, Nigeria v1 First published: 20 Sep 2018, 7:1504 Open Peer Review https://doi.org/10.12688/f1000research.16058.1 Latest published: 20 Sep 2018, 7:1504 https://doi.org/10.12688/f1000research.16058.1 Reviewer Status Invited Reviewers Abstract Background: Heat shock proteins (HSPs) are molecular chaperones 1 2 3 known to bind and sequester client proteins under stress. Methods: To identify and better understand some of these proteins, version 1 we carried out a computational genome-wide survey of the bovine 20 Sep 2018 report report report genome.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • The Endoplasmic Reticulum Chaperone Erdj4 Is Required for Survival, Glucose Metabolism and B Cell Development
    The endoplasmic reticulum chaperone ERdj4 is required for survival, glucose metabolism and B cell development A dissertation submitted to the University of Cincinnati in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY in the Immunobiology Graduate Program of the College of Medicine 2012 by Jill Marie Fritz B.A., Miami University, Oxford, OH Advisory Committee: Timothy Weaver, M.S., Ph.D., Chair George Deepe, M.D. Fred Finkelman, M.D. H. Leighton Grimes, Ph.D. Christopher Karp, M.D. Francis McCormack, M.D. ABSTRACT The ER-localized DnaJ homologue 4 (ERdj4) is a soluble ER chaperone induced by the unfolded protein response (UPR) to assist in the removal of unfolded/misfolded proteins from the ER lumen for proteasomal degradation. To elucidate the function of ERdj4 in vivo, ERdj4 gene trap (ERdj4GT/GT) mice were generated from embryonic stem cells harboring a gene trap cassette inserted into the ERdj4 locus. ERdj4GT/GT mice expressed hypomorphic levels of ERdj4 with a 10-100 fold reduction in all tissues and cell types examined. Approximately 30-50% of ERdj4GT/GT mice died perinatally in association with growth retardation and hypoglycemia. ERdj4GT/GT neonates exhibited signs of delayed pancreatic development, including abnormal distribution of pancreatic α- and β-cells and reduced insulin and glucagon in the pancreas. Surviving adult ERdj4GT/GT mice were glucose intolerant, resulting from hypoinsulinemia rather than insulin resistance. Pancreatic β-cells exhibited increased ER stress, including ER dilation and upregulation of the UPR. Proinsulin accumulated in the ER of β-cells from ERdj4GT/GT mice consistent with our previous finding that proinsulin is a substrate for ERdj4.
    [Show full text]
  • Pdf 568.24 K
    Biomacromol. J. Vol. 2, No. 2, 169-176, December 2016. Biomacromolecular Journal www.bmmj.org Dendrosomal Nanocurcumin Induces Changes in the Expression Levels of Heat Shock Proteins in the AGS Cell Line: Cellular Tolerance or Smart Apoptosis Induction A. Panahia,e, R. Nakhaei Sistanib,*, M. Entezaric, H. Shirzada, M. Montazerid, S. Babashaha and M. Sadeghizadeha,* aDepartment of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran bFaculty of Chemistry, Department of Biotechnology, University of Kashan, Kashan, Iran cDepartment of Genetics, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran dDepartment of Medical Biotechnology, Tehran Medical Sciences Branch, Islamic Azad University, Tehran, Iran eDepartment of Biology, Faculty of Basic Sciences, University of Mohaghegh Ardabili , Ardabil , Iran (Received 8 March 2017, Accepted 13 May 2017) ABSTRACT Objectives: The expression levels of heat shock proteins (HSPs) are elevated in many cancers, and this overexpression is often associated with both a poor survival and a therapeutic outcome. Curcumin is an anti-cancer agent that also induces a heat shock response. HSPs confer resistance to curcumin-induced apoptosis in cancerous cells. The aim of the current study was to analyze variations in the expression levels of hsp gene in response to polymeric-nanocurcumin (DNC) to determine whether these alterations have inhibitory effects on apoptosis or promote it. Methods: The IC50 of DNC on AGS cell line was determined by the MTT assay. Following DNC treatment, the expression levels of hsp mRNAs were quantified by qPCR. Using siRNA, the Hsp27 gene was knocked down, and the DNC-induced apoptosis was determined by Annexin V test.
    [Show full text]
  • The HSP70 Chaperone Machinery: J Proteins As Drivers of Functional Specificity
    REVIEWS The HSP70 chaperone machinery: J proteins as drivers of functional specificity Harm H. Kampinga* and Elizabeth A. Craig‡ Abstract | Heat shock 70 kDa proteins (HSP70s) are ubiquitous molecular chaperones that function in a myriad of biological processes, modulating polypeptide folding, degradation and translocation across membranes, and protein–protein interactions. This multitude of roles is not easily reconciled with the universality of the activity of HSP70s in ATP-dependent client protein-binding and release cycles. Much of the functional diversity of the HSP70s is driven by a diverse class of cofactors: J proteins. Often, multiple J proteins function with a single HSP70. Some target HSP70 activity to clients at precise locations in cells and others bind client proteins directly, thereby delivering specific clients to HSP70 and directly determining their fate. In their native cellular environment, polypeptides are participates in such diverse cellular functions. Their constantly at risk of attaining conformations that pre- functional diversity is remarkable considering that vent them from functioning properly and/or cause them within and across species, HSP70s have high sequence to aggregate into large, potentially cytotoxic complexes. identity. They share a single biochemical activity: an Molecular chaperones guide the conformation of proteins ATP-dependent client-binding and release cycle com- throughout their lifetime, preventing their aggregation bined with client protein recognition, which is typi- by protecting interactive surfaces against non-productive cally rather promiscuous. This apparent conundrum interactions. Through such inter actions, molecular chap- is resolved by the fact that HSP70s do not work alone, erones aid in the folding of nascent proteins as they are but rather as ‘HSP70 machines’, collaborating with synthesized by ribosomes, drive protein transport across and being regulated by several cofactors.
    [Show full text]
  • Senescence Inhibits the Chaperone Response to Thermal Stress
    SUPPLEMENTAL INFORMATION Senescence inhibits the chaperone response to thermal stress Jack Llewellyn1, 2, Venkatesh Mallikarjun1, 2, 3, Ellen Appleton1, 2, Maria Osipova1, 2, Hamish TJ Gilbert1, 2, Stephen M Richardson2, Simon J Hubbard4, 5 and Joe Swift1, 2, 5 (1) Wellcome Centre for Cell-Matrix Research, Oxford Road, Manchester, M13 9PT, UK. (2) Division of Cell Matrix Biology and Regenerative Medicine, School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, University of Manchester, Manchester, M13 9PL, UK. (3) Current address: Department of Biomedical Engineering, University of Virginia, Box 800759, Health System, Charlottesville, VA, 22903, USA. (4) Division of Evolution and Genomic Sciences, School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, University of Manchester, Manchester, M13 9PL, UK. (5) Correspondence to SJH ([email protected]) or JS ([email protected]). Page 1 of 11 Supplemental Information: Llewellyn et al. Chaperone stress response in senescence CONTENTS Supplemental figures S1 – S5 … … … … … … … … 3 Supplemental table S6 … … … … … … … … 10 Supplemental references … … … … … … … … 11 Page 2 of 11 Supplemental Information: Llewellyn et al. Chaperone stress response in senescence SUPPLEMENTAL FIGURES Figure S1. A EP (passage 3) LP (passage 16) 200 µm 200 µm 1.5 3 B Mass spectrometry proteomics (n = 4) C mRNA (n = 4) D 100k EP 1.0 2 p < 0.0001 p < 0.0001 LP p < 0.0001 p < 0.0001 ) 0.5 1 2 p < 0.0001 p < 0.0001 10k 0.0 0 -0.5 -1 Cell area (µm Cell area fold change vs. EP fold change vs.
    [Show full text]
  • Molecular Signatures Differentiate Immune States in Type 1 Diabetes Families
    Page 1 of 65 Diabetes Molecular signatures differentiate immune states in Type 1 diabetes families Yi-Guang Chen1, Susanne M. Cabrera1, Shuang Jia1, Mary L. Kaldunski1, Joanna Kramer1, Sami Cheong2, Rhonda Geoffrey1, Mark F. Roethle1, Jeffrey E. Woodliff3, Carla J. Greenbaum4, Xujing Wang5, and Martin J. Hessner1 1The Max McGee National Research Center for Juvenile Diabetes, Children's Research Institute of Children's Hospital of Wisconsin, and Department of Pediatrics at the Medical College of Wisconsin Milwaukee, WI 53226, USA. 2The Department of Mathematical Sciences, University of Wisconsin-Milwaukee, Milwaukee, WI 53211, USA. 3Flow Cytometry & Cell Separation Facility, Bindley Bioscience Center, Purdue University, West Lafayette, IN 47907, USA. 4Diabetes Research Program, Benaroya Research Institute, Seattle, WA, 98101, USA. 5Systems Biology Center, the National Heart, Lung, and Blood Institute, the National Institutes of Health, Bethesda, MD 20824, USA. Corresponding author: Martin J. Hessner, Ph.D., The Department of Pediatrics, The Medical College of Wisconsin, Milwaukee, WI 53226, USA Tel: 011-1-414-955-4496; Fax: 011-1-414-955-6663; E-mail: [email protected]. Running title: Innate Inflammation in T1D Families Word count: 3999 Number of Tables: 1 Number of Figures: 7 1 For Peer Review Only Diabetes Publish Ahead of Print, published online April 23, 2014 Diabetes Page 2 of 65 ABSTRACT Mechanisms associated with Type 1 diabetes (T1D) development remain incompletely defined. Employing a sensitive array-based bioassay where patient plasma is used to induce transcriptional responses in healthy leukocytes, we previously reported disease-specific, partially IL-1 dependent, signatures associated with pre and recent onset (RO) T1D relative to unrelated healthy controls (uHC).
    [Show full text]
  • Roles of Xbp1s in Transcriptional Regulation of Target Genes
    biomedicines Review Roles of XBP1s in Transcriptional Regulation of Target Genes Sung-Min Park , Tae-Il Kang and Jae-Seon So * Department of Medical Biotechnology, Dongguk University, Gyeongju 38066, Gyeongbuk, Korea; [email protected] (S.-M.P.); [email protected] (T.-I.K.) * Correspondence: [email protected] Abstract: The spliced form of X-box binding protein 1 (XBP1s) is an active transcription factor that plays a vital role in the unfolded protein response (UPR). Under endoplasmic reticulum (ER) stress, unspliced Xbp1 mRNA is cleaved by the activated stress sensor IRE1α and converted to the mature form encoding spliced XBP1 (XBP1s). Translated XBP1s migrates to the nucleus and regulates the transcriptional programs of UPR target genes encoding ER molecular chaperones, folding enzymes, and ER-associated protein degradation (ERAD) components to decrease ER stress. Moreover, studies have shown that XBP1s regulates the transcription of diverse genes that are involved in lipid and glucose metabolism and immune responses. Therefore, XBP1s has been considered an important therapeutic target in studying various diseases, including cancer, diabetes, and autoimmune and inflammatory diseases. XBP1s is involved in several unique mechanisms to regulate the transcription of different target genes by interacting with other proteins to modulate their activity. Although recent studies discovered numerous target genes of XBP1s via genome-wide analyses, how XBP1s regulates their transcription remains unclear. This review discusses the roles of XBP1s in target genes transcriptional regulation. More in-depth knowledge of XBP1s target genes and transcriptional regulatory mechanisms in the future will help develop new therapeutic targets for each disease. Citation: Park, S.-M.; Kang, T.-I.; Keywords: XBP1s; IRE1; ATF6; ER stress; unfolded protein response; UPR; RIDD So, J.-S.
    [Show full text]
  • Prognostic and Functional Significant of Heat Shock Proteins (Hsps)
    biology Article Prognostic and Functional Significant of Heat Shock Proteins (HSPs) in Breast Cancer Unveiled by Multi-Omics Approaches Miriam Buttacavoli 1,†, Gianluca Di Cara 1,†, Cesare D’Amico 1, Fabiana Geraci 1 , Ida Pucci-Minafra 2, Salvatore Feo 1 and Patrizia Cancemi 1,2,* 1 Department of Biological Chemical and Pharmaceutical Sciences and Technologies (STEBICEF), University of Palermo, 90128 Palermo, Italy; [email protected] (M.B.); [email protected] (G.D.C.); [email protected] (C.D.); [email protected] (F.G.); [email protected] (S.F.) 2 Experimental Center of Onco Biology (COBS), 90145 Palermo, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-091-2389-7330 † These authors contributed equally to this work. Simple Summary: In this study, we investigated the expression pattern and prognostic significance of the heat shock proteins (HSPs) family members in breast cancer (BC) by using several bioinfor- matics tools and proteomics investigations. Our results demonstrated that, collectively, HSPs were deregulated in BC, acting as both oncogene and onco-suppressor genes. In particular, two different HSP-clusters were significantly associated with a poor or good prognosis. Interestingly, the HSPs deregulation impacted gene expression and miRNAs regulation that, in turn, affected important bio- logical pathways involved in cell cycle, DNA replication, and receptors-mediated signaling. Finally, the proteomic identification of several HSPs members and isoforms revealed much more complexity Citation: Buttacavoli, M.; Di Cara, of HSPs roles in BC and showed that their expression is quite variable among patients. In conclusion, G.; D’Amico, C.; Geraci, F.; we elaborated two panels of HSPs that could be further explored as potential biomarkers for BC Pucci-Minafra, I.; Feo, S.; Cancemi, P.
    [Show full text]
  • Characterization of the Dual Functional Effects of Heat Shock Proteins (Hsps
    Zhang et al. Genome Medicine (2020) 12:101 https://doi.org/10.1186/s13073-020-00795-6 RESEARCH Open Access Characterization of the dual functional effects of heat shock proteins (HSPs) in cancer hallmarks to aid development of HSP inhibitors Zhao Zhang1†, Ji Jing2†, Youqiong Ye1, Zhiao Chen1, Ying Jing1, Shengli Li1, Wei Hong1, Hang Ruan1, Yaoming Liu1, Qingsong Hu3, Jun Wang4, Wenbo Li1, Chunru Lin3, Lixia Diao5*, Yubin Zhou2* and Leng Han1* Abstract Background: Heat shock proteins (HSPs), a representative family of chaperone genes, play crucial roles in malignant progression and are pursued as attractive anti-cancer therapeutic targets. Despite tremendous efforts to develop anti-cancer drugs based on HSPs, no HSP inhibitors have thus far reached the milestone of FDA approval. There remains an unmet need to further understand the functional roles of HSPs in cancer. Methods: We constructed the network for HSPs across ~ 10,000 tumor samples from The Cancer Genome Atlas (TCGA) and ~ 10,000 normal samples from Genotype-Tissue Expression (GTEx), and compared the network disruption between tumor and normal samples. We then examined the associations between HSPs and cancer hallmarks and validated these associations from multiple independent high-throughput functional screens, including Project Achilles and DRIVE. Finally, we experimentally characterized the dual function effects of HSPs in tumor proliferation and metastasis. Results: We comprehensively analyzed the HSP expression landscape across multiple human cancers and revealed a global disruption of the co-expression network for HSPs. Through analyzing HSP expression alteration and its association with tumor proliferation and metastasis, we revealed dual functional effects of HSPs, in that they can simultaneously influence proliferation and metastasis in opposite directions.
    [Show full text]
  • Protein Folding and Quality Control in the ER
    Downloaded from http://cshperspectives.cshlp.org/ on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press Protein Folding and Quality Control in the ER Kazutaka Araki and Kazuhiro Nagata Laboratory of Molecular and Cellular Biology, Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo, Kita-ku, Kyoto 803-8555, Japan Correspondence: [email protected] The endoplasmic reticulum (ER) uses an elaborate surveillance system called the ER quality control (ERQC) system. The ERQC facilitates folding and modification of secretory and mem- brane proteins and eliminates terminally misfolded polypeptides through ER-associated degradation (ERAD) or autophagic degradation. This mechanism of ER protein surveillance is closely linked to redox and calcium homeostasis in the ER, whose balance is presumed to be regulated by a specific cellular compartment. The potential to modulate proteostasis and metabolism with chemical compounds or targeted siRNAs may offer an ideal option for the treatment of disease. he endoplasmic reticulum (ER) serves as a complex in the ER membrane (Johnson and Tprotein-folding factory where elaborate Van Waes 1999; Saraogi and Shan 2011). After quality and quantity control systems monitor arriving at the translocon, translation resumes an efficient and accurate production of secretory in a process called cotranslational translocation and membrane proteins, and constantly main- (Hegde and Kang 2008; Zimmermann et al. tain proper physiological homeostasis in the 2010). Numerous ER-resident chaperones and ER including redox state and calcium balance. enzymes aid in structural and conformational In this article, we present an overview the recent maturation necessary for proper protein fold- progress on the ER quality control system, ing, including signal-peptide cleavage, N-linked mainly focusing on the mammalian system.
    [Show full text]