DOCSLIB.ORG

Receptor for Hyaluronan-Mediated Motility Isoform B Promotes Liver Metastasis in a Mouse Model of Multistep Tumorigenesis and a Tail Vein Assay for Metastasis

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Receptor for Hyaluronan-Mediated Motility Isoform B Promotes Liver Metastasis in a Mouse Model of Multistep Tumorigenesis and a Tail Vein Assay for Metastasis Receptor for hyaluronan-mediated motility isoform B promotes liver metastasis in a mouse model of multistep tumorigenesis and a tail vein assay for metastasis Yi-Chieh Nancy Dua,b,1, Chen-Kung Chouc, David S. Klimstrad, and Harold Varmusa,e,1 aProgram in Cancer Biology and Genetics, Memorial Sloan-Kettering Cancer Center, New York, NY 10065; bDepartment of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY 10065; cDepartment of Biomedical Sciences, Chang-Gung University, Tao-Yuan 333, Taiwan; dDepartment of Pathology, Memorial Sloan-Kettering Cancer Center, New York, NY 10065; and eCancer Biology and Genetics Section, Cancer Genetics Branch, National Human Genome Research Institute, Bethesda, MD 20892 Contributed by Harold Varmus, August 30, 2011 (sent for review July 14, 2011) The gene encoding the receptor for hyaluronan-mediated motility Using the RIP-Tag; RIP-tva model, we find that isoform B of B (RHAMM) is overexpressed in many human cancers. However, it RHAMM (RHAMM ) enhances the growth of mouse islet is unclear whether RHAMM plays a causal role in tumor initiation tumors and promotes metastasis exclusively to the liver and local or progression. Using somatic gene transfer in a mouse model of lymph nodes. Furthermore, we show that mouse pancreatic islet B islet cell tumorigenesis, we demonstrate that RHAMM isoform B tumor cells programmed to express RHAMM form hepatic me- (RHAMMB) promotes tumor growth and metastases to lymph tastasis when injected into the tail vein of mice in a traditional nodes and the liver. The propensity of RHAMMB-expressing cells assay for metastasis. The cells also show evidence that RHAMM fi has enhanced signaling via the epidermal growth factor receptor to metastasize to the liver was con rmed using an experimental B metastasis assay in which cells were injected into the tail vein of (EGFR). These observations and others suggest that RHAMM B may be an important factor in tumor growth and progression and immunodeficient mice. However, RHAMM did not increase cell that a better understanding of the RHAMM gene might offer migration or proliferation in culture. In initial efforts to identify insights into the organotropism of metastatic cancer cells. signaling pathways activated by RHAMMB, we found that B RHAMM induced phosphorylation of epidermal growth factor re- Results ceptor (EGFR), Erk1/2, and STAT3 and conferred susceptibility to RHAMMB Promotes Tumor Growth and Metastasis to Pancreatic apoptosis after treatment with an EGFR inhibitor, gefitinib. Taken Lymph Nodes and the Liver in a Mouse Model of Pancreatic Islet together, the results indicate that RHAMMB promotes hepatic Tumors. To evaluate the malignant potential of genes reported metastasis by islet tumor cells, perhaps through growth factor to be up-regulated both in HCC and during mouse liver re- receptor-mediated signaling. generation (2), we examined five candidate genes, including RHAMMB, paternally expressed 10 (PEG10), FLJ10540, FLJ11252, e have previously reported a bitransgenic mouse model, and FLJ11164. FLJ stands for the “full-length long Japan” col- WRIP-Tag; RIP-tva, in which the rat insulin promoter (RIP) lection of human cDNAs (8). drives production of both the SV40 T antigen (RIP-Tag) and The cDNAs of the candidates were cloned into avian retroviral the receptor for subgroup A avian leukosis virus (RIP-tva)in vector, RCASBP, with a FLAG epitope tag added to the N pancreatic β cells (1). Coding domains of genes suspected of terminus of PEG10, FLJ10540, and FLJ11252. We injected high 8 contributing to tumor progression can be introduced into pre- titer virus stocks (0.1 mL; >10 infectious units per milliliter) into malignant lesions by infection with the avian retroviral vector, 7-wk-old RIP-Tag; RIP-tva mice by the intracardiac route. At this RCASBP, after intracardiac injection. RIP-Tag transgenic mice point, many islets show evidence of hyperplasia, allowing in- develop islet tumors through well-defined stages that resemble fection with oncoretrovirus vectors, which are dependent on – the progression of several kinds of human cancers; for this rea- cell division for successful infection (1). RCASBP ALPP (Al- MEDICAL SCIENCES – son, we and others have used these mice, with or without addi- kaline Phosphatase) and RCASBP Bcl-xL were chosen as con- tional transgenes, to identify and validate mechanisms of trols. ALPP encodes a protein unlikely to contribute to tumori- genesis, serving as a negative control for effects of viral infection. tumorigenesis that may operate in multiple tissues. For instance, – we have used RIP-Tag; RIP-tva mice to show that RCASBP- We have previously shown that infection with RCASBP Bcl-xL promotes tumor growth and lymph node metastasis in RIP-Tag; mediated delivery of Bcl-xL or E-cadherin, factors implicated in RIP-tva mice (1), so infection with this virus provided a pos- various neoplasms, promotes tumorigenesis and invasion in islet itive control. cells (1). fi Nine weeks after infection, the pancreas and other organs High-throughput genomic technologies have identi ed many were harvested for histological staging and grading of the lesions. genes that may be critical in tumor initiation and progression. Human RHAMMB significantly increased pancreatic tumor However, it remains difficult to distinguish causative and pas- fi burden in 8 of 12 mice, but not all, compared with mice infected senger mutations and to assign speci c biological functions to with RCASBP–ALPP (Fig. 1, P = 0.0097, Wilcoxon rank sum altered genes in human cancers, and the RIP-Tag; RIP-tva mouse test). RCASBP–Bcl-xL induced a small increase in pancreatic model of multistage tumorigenesis offers an opportunity to ad- tumor burden (P = 0.0087), whereas none of the other vec- dress such issues. To that end, we have assessed the oncogenic functions of a small number of incompletely characterized genes that are up-regulated in human hepatocellular carcinomas Author contributions: Y.-C.N.D. and H.V. designed research; Y.-C.N.D. performed re- (HCC) and during mouse liver regeneration (2). One of the search; C.-K.C. provided new reagents; D.S.K. contributed pathology analysis; Y.-C.N.D. candidate genes, a gene encoding a receptor for hyaluronan- analyzed data; and Y.-C.N.D. and H.V. wrote the paper. mediated motility (RHAMM) is overexpressed in many types of The authors declare no conflict of interest. human cancers, including pancreatic ductal carcinoma, hepato- Freely available online through the PNAS open access option. cellular carcinoma, multiple myeloma, breast cancer, gliomas, 1To whom correspondence may be addressed. E-mail: [email protected] or colon cancer, and prostate cancer (2–7); but the functions of at [email protected]. least four proteins encoded by its alternatively spliced messenger This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. RNAs and their roles, if any, in tumorigenesis are unclear. 1073/pnas.1114022108/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1114022108 PNAS | October 4, 2011 | vol. 108 | no. 40 | 16753–16758 Downloaded by guest on September 28, 2021 tors (RCASBP–FLAG-PEG10, RCASBP–FLAG-FLJ10540, Table 1. Impact of candidate genes on tumorigenesis in vivo RCASBP–FLAG-FLJ11252, or RCASBP–FLJ11164) caused fi Tumor any signi cant increase in tumor burden (Table 1 and Table S1). Age burden Lymph node Liver To aid the search for metastasis of islet tumors in RIP-Tag; RCASBP– (wk) (mm3) metastasis (%) metastasis (%) RIP-tva mice, tissue sections were subjected to immunohisto- chemical staining for synaptophysin, a neuroendocrine marker, ALPP 12 8.1 ± 4.5 0/5 mice (0) 0/5 mice (0) and for insulin, a β-cell marker. Local lymph node metastases ALPP 16 99.7 ± 19.4 0/10 mice (0) 0/10 mice (0) – B were detected in mice infected with RCASBP RHAMM (8 of RHAMMB 12 16.2 ± 8.9 0/6 mice (0) 0/6 mice (0) 11 mice, P = 0.001, Fisher’s exact test), RCASBP–Bcl-xL (7 of 15 B ± – RHAMM 16 298.3 121.3 8/11 mice (73) 8/12 mice (67) mice, P = 0.013), RCASBP FLAG-PEG10 (2 of 8 mice, P = Bcl-xL 16 150.5 ± 23 7/15 mice (46) 0/6 mice (0) 0.183), RCASBP–FLAG-FLJ10540 (4 of 7 mice, P = 0.015), and ± – FLAG-PEG10 16 57.8 26.8 2/8 mice (25) 0/8 mice (0) RCASBP FLJ11164 (1 of 9 mice, P = 0.474) (Fig. 2, Table 1, FLAG-FLJ10540 16 126.1 ± 54.3 4/7 mice (57) 2/7 mice (28) and Table S1). The sizes of lymph node metastases in mice – – FLAG-FLJ11252 16 41.2 ± 30.9 0/7 mice (0) 0/7 mice (0) infected with RCASBP FLAG-PEG10,RCASBPFLAG- ± FLJ10540, and RCASBP–FLJ11164 were small and were not FLJ11164 16 43.8 25.1 1/9 mice (11) 0/6 mice (0)* easily detected by hematoxylin and eosin staining, unlike those in RCASBP retroviruses carrying indicated cDNAs were introduced to RIP- B mice infected with RCASBP–RHAMM and RCASBP–Bcl-xL. Tag; RIP-tva mice through intracardiac injection at 7 wk of age. Mice were No lymph node metastases were found in mice receiving the euthanized at 12 or 16 wk of age for measurement of tumor burden and for negative control virus or RCASBP–FLAG-FLJ11252 (Table 1 metastasis survey. A standard formula for tumor volume was applied (vol- and Table S1). ume [mm3] = 0.52 × width2 × length). Tumor burden is the sum of the tumor Importantly, liver metastases were found in 8 of 12 RCASBP– volume per mouse. RHAMMB infected mice (Fig. 2, Table 1, and Table S1, P = *Mice having micrometastases with fewer than five cells were excluded. 0.002, Fisher’s exact test), and the appearance of metastasis was not closely associated with the aggregated size of primary tumors.
Load more

Recommended publications
  • Loss of BCL-3 Sensitises Colorectal Cancer Cells to DNA Damage, Revealing A
    bioRxiv preprint doi: https://doi.org/10.1101/2021.08.03.454995; this version posted August 4, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Title: Loss of BCL-3 sensitises colorectal cancer cells to DNA damage, revealing a role for BCL-3 in double strand break repair by homologous recombination Authors: Christopher Parker*1, Adam C Chambers*1, Dustin Flanagan2, Tracey J Collard1, Greg Ngo3, Duncan M Baird3, Penny Timms1, Rhys G Morgan4, Owen Sansom2 and Ann C Williams1. *Joint first authors. Author affiliations: 1. Colorectal Tumour Biology Group, School of Cellular and Molecular Medicine, Faculty of Life Sciences, Biomedical Sciences Building, University Walk, University of Bristol, Bristol, BS8 1TD, UK 2. Cancer Research UK Beatson Institute, Garscube Estate, Switchback Road, Bearsden Glasgow, G61 1BD UK 3. Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, CF14 4XN UK 4. School of Life Sciences, University of Sussex, Sussex House, Falmer, Brighton, BN1 9RH UK 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.08.03.454995; this version posted August 4, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract (250 words) Objective: The proto-oncogene BCL-3 is upregulated in a subset of colorectal cancers (CRC) and increased expression of the gene correlates with poor patient prognosis. The aim is to investigate whether inhibiting BCL-3 can increase the response to DNA damage in CRC.
    [Show full text]
  • Identification and Characterization of Missense Alterations in the BRCA1
    633 ORIGINAL ARTICLE Identification and characterization of missense alterations in the BRCA1 associated RING domain (BARD1) gene in breast and ovarian cancer M K Sauer, I L Andrulis ............................................................................................................................... J Med Genet 2005;42:633–638. doi: 10.1136/jmg.2004.030049 Background: BRCA1 associated RING domain protein (BARD1) was originally identified due to its interaction with the RING domain of BRCA1. BARD1 is required for S phase progression, contact inhibition and normal nuclear division, as well as for BRCA1 independent, p53 dependent apoptosis. See end of article for Methods: To investigate whether alterations in BARD1 are involved in human breast and ovarian cancer, authors’ affiliations ....................... we used single strand conformation polymorphism analysis and sequencing on 35 breast tumours and cancer cell lines and on 21 ovarian tumours. Correspondence to: Results: Along with the G2355C (S761N) missense mutation previously identified in a uterine cancer, we Dr M K Sauer, Samuel Lunenfeld Research found two other variants in breast cancers, T2006C (C645R) and A2286G (I738V). The T2006C (C645R) Institute, Mount Sinai mutation was also found in one ovarian tumour. A variant of uncertain consequence, G1743C (C557S), Hospital, 600 University was found to be homozygous or hemizygous in an ovarian tumour. Eleven variants of BARD1 were Ave, Toronto, Ontario, M5G 1X5, Canada; characterised with respect to known functions of BARD1. None of the variants appears to affect [email protected] localisation or interaction with BRCA1; however, putative disease associated alleles appear to affect the stability of p53. These same mutations also appear to abrogate the growth suppressive and apoptotic Received activities of BARD1.
    [Show full text]
  • Functional Characterization of Full-Length BARD1 Strengthens Its
    Journal of Cancer 2020, Vol. 11 1495 Ivyspring International Publisher Journal of Cancer 2020; 11(6): 1495-1504. doi: 10.7150/jca.36164 Research Paper Functional characterization of full-length BARD1 strengthens its role as a tumor suppressor in neuroblastoma Flora Cimmino1,2, Marianna Avitabile1,2, Vito Alessandro Lasorsa1,2, Lucia Pezone1, Antonella Cardinale1, Annalaura Montella2, Sueva Cantalupo3, Achille Iolascon1,2, Mario Capasso1,2,3 1. Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli “Federico II”, Naples, Italy 2. CEINGE Biotecnologie Avanzate, Naples, Italy 3. IRCCS SDN, Naples, Italy Corresponding author: Flora Cimmino, phD, University of Naples Federico II, Department of Molecular Medicine and Medical Biotechnology, CEINGE Biotecnologie Avanzate, Via Gaetano Salvatore, 486, 80145 Napoli Italy. Lab: +39 0813737736; Fax: +39 0813737804; Email: [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: 2019.04.29; Accepted: 2019.11.12; Published: 2020.01.14 Abstract BARD1 is associated with the development of high-risk neuroblastoma patients. Particularly, the expression of full length (FL) isoform, FL BARD1, correlates to high-risk neuroblastoma development and its inhibition is sufficient to induce neuroblastoma cells towards a worst phenotype. Here we have investigated the mechanisms of FL BARD1 in neuroblastoma cell lines depleted for FL BARD1 expression. We have shown that FL BARD1 expression protects the cells from spontaneous DNA damage and from damage accumulated after irradiation. We demonstrated a role for FL BARD1 as tumor suppressor to prevent unscheduled mitotic entry of DNA damaged cells and to lead to death cells that have bypassed cell cycle checkpoints.
    [Show full text]
  • BRCA1–BARD1 Regulates Axon Regeneration in Concert with the Gqα–DAG Signaling Network
    Research Articles: Cellular/Molecular BRCA1–BARD1 regulates axon regeneration in concert with the Gqα–DAG signaling network https://doi.org/10.1523/JNEUROSCI.1806-20.2021 Cite as: J. Neurosci 2021; 10.1523/JNEUROSCI.1806-20.2021 Received: 13 July 2020 Revised: 20 January 2021 Accepted: 5 February 2021 This Early Release article has been peer-reviewed and accepted, but has not been through the composition and copyediting processes. The final version may differ slightly in style or formatting and will contain links to any extended data. Alerts: Sign up at www.jneurosci.org/alerts to receive customized email alerts when the fully formatted version of this article is published. Copyright © 2021 Sakai et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed. Sakai et al., 1 1 BRCA1–BARD1 regulates axon regeneration in 2 concert with the GqD–DAG signaling network 3 4 Abbreviated Title: BRCA1–BARD1 regulates axon regeneration 5 6 Yoshiki Sakai, Hiroshi Hanafusa, Tatsuhiro Shimizu, Strahil Iv. 7 Pastuhov, Naoki Hisamoto, and Kunihiro Matsumoto 8 9 Division of Biological Science, Graduate School of Science, Nagoya University, 10 Chikusa-ku, Nagoya 464-8602, Japan 11 12 To whom correspondence should be addressed: 13 Kunihiro Matsumoto and Naoki Hisamoto 14 Division of Biological Science, Graduate School of Science, 15 Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan. 16 E-mail address: [email protected] (K.
    [Show full text]
  • BARD1 in Cell Life and Death
    Role of BARD1 in cell life and death Irmgard Irminger-Finger Biology of Aging Laboratory Department of Geriatrics University of Geneva Switzerland Age is the biggest risk factor for cancer DePinho, 2000 Cancer Risks for Men prostate From1950 1990 Cancer Risks for Women breast From1950 1990 Cancers of old age are different from young age cancers Cancer in old age has a different face than cancer in young age BCC Basal cell carcinoma SCC Squamous cell carcinoma DePinho, 2000 What could link epithelial cell derived cancers to aging? BARD1, not just another cancer predisposition gene Common pathways for cancer and aging? Accumulation of damage DNA damage DNA damage proliferation- DNA damage arrest repair senescence proliferation cancer elimination apoptosis Cellular aging BARD1 Summary • BARD1 structure • BARD1 repression studies • BARD1 overexpression studies • BARD1 dynamic localization • BARD1 induced upon stress • Non-correlated expression of BARD1 and BRCA1 • Role in spermatogenesis • BARD1 upregulation upon stress • Role in tumorigenesis • BARD1 a tumor antigen • BARD1 in cancer vaccine and genetherapy Conserved structures in BARD1 and BRCA1 Q>H RING ANK BRCT BARD1 66 95 53 97 77 91 conservation RING BRCT BRCA1 C>G No one like BARD1 RING ANK BRCT BARD1 BRCA1 Arabidopsis 2 hypoth.p. Arabidopsis CAC05430 C. elegans 3 hypoth. p. C. elegans 6 hypoth. p. C. elegans T15564 C. elegans T15566 Xenopus BARD1 Joukov, Livingston, PNAS, 2001 BRCA1 BARD1- BRCA1 heterodimer Rad51 PCNA ? RNA-Pol II p53 BAP1 Nuclear localization BACH1 RHA signals Granin BRCA2 motif BARD1 interaction Ankyrin repeats BRCT BRCA1 domain interaction Ubiquitin-ligase Polyadenylation activity A B C D BARD1 BRCA1 BARD1 BRCA1 BARD1 Bcl-3 Pol II/Holo CstF-50 NF-kB Replication? S-phase dots Transcription: mRNA Regulation of Pol II/Holo Interaction Processing? transcription? [Scully et al.
    [Show full text]
  • Hyaluronan Mediated Motility Receptor (HMMR) Encodes an Evolutionarily Conserved Homeostasis, Mitosis, and Meiosis Regulator Rather Than a Hyaluronan Receptor
    cells Review Hyaluronan Mediated Motility Receptor (HMMR) Encodes an Evolutionarily Conserved Homeostasis, Mitosis, and Meiosis Regulator Rather than a Hyaluronan Receptor 1 1 1, 1,2, Zhengcheng He , Lin Mei , Marisa Connell y and Christopher A. Maxwell * 1 Department of Pediatrics, University of British Columbia, Vancouver, BC V5Z 4H4, Canada; [email protected] (Z.H.); [email protected] (L.M.); [email protected] (M.C.) 2 Michael Cuccione Childhood Cancer Research Program, BC Children’s Hospital, Vancouver, BC V5Z 4H4, Canada * Correspondence: [email protected]; Tel.: +1-6048752000 (ext. 4691) Current position: Department of Neuroscience and Physiology, SUNY Upstate Medical University, Syracuse, y NY 13210, USA. Received: 3 March 2020; Accepted: 25 March 2020; Published: 28 March 2020 Abstract: Hyaluronan is an extracellular matrix component that absorbs water in tissues and engages cell surface receptors, like Cluster of Differentiation 44 (CD44), to promote cellular growth and movement. Consequently, CD44 demarks stem cells in normal tissues and tumor-initiating cells isolated from neoplastic tissues. Hyaluronan mediated motility receptor (HMMR, also known as RHAMM) is another one of few defined hyaluronan receptors. HMMR is also associated with neoplastic processes and its role in cancer progression is often attributed to hyaluronan-mediated signaling. But, HMMR is an intracellular, microtubule-associated, spindle assembly factor that localizes protein complexes to augment the activities of mitotic kinases, like polo-like kinase 1 and Aurora kinase A, and control dynein and kinesin motor activities. Expression of HMMR is elevated in cells prior to and during mitosis and tissues with detectable HMMR expression tend to be highly proliferative, including neoplastic tissues.
    [Show full text]
  • Isomerization of BRCA1-BARD1 Promotes Replication Fork Protection
    University of Birmingham Isomerization of BRCA1-BARD1 promotes replication fork protection Daza-Martin, Manuel; Starowicz, Katarzyna; Jamshad, Mohammed; Tye, Stephanie; Ronson, George E; MacKay, Hannah L; Chauhan, Anoop Singh; Walker, Alexandra K; Stone, Helen R; Beesley, James F J; Coles, Jennifer L; Garvin, Alexander J; Stewart, Grant S; McCorvie, Thomas J; Zhang, Xiaodong; Densham, Ruth M; Morris, Joanna R DOI: 10.1038/s41586-019-1363-4 License: None: All rights reserved Document Version Peer reviewed version Citation for published version (Harvard): Daza-Martin, M, Starowicz, K, Jamshad, M, Tye, S, Ronson, GE, MacKay, HL, Chauhan, AS, Walker, AK, Stone, HR, Beesley, JFJ, Coles, JL, Garvin, AJ, Stewart, GS, McCorvie, TJ, Zhang, X, Densham, RM & Morris, JR 2019, 'Isomerization of BRCA1-BARD1 promotes replication fork protection', Nature, vol. 571, no. 7766, pp. 521-527. https://doi.org/10.1038/s41586-019-1363-4 Link to publication on Research at Birmingham portal Publisher Rights Statement: Checked for eligibility: 11/07/2019 https://www.nature.com/articles/s41586-019-1363-4 Daza-Martin, Manuel, et al. "Isomerization of BRCA1–BARD1 promotes replication fork protection." Nature (2019): 1. https://doi.org/10.1038/s41586-019-1363-4 General rights Unless a licence is specified above, all rights (including copyright and moral rights) in this document are retained by the authors and/or the copyright holders. The express permission of the copyright holder must be obtained for any use of this material other than for purposes permitted by law. •Users may freely distribute the URL that is used to identify this publication. •Users may download and/or print one copy of the publication from the University of Birmingham research portal for the purpose of private study or non-commercial research.
    [Show full text]
  • BRCC3 Acts As a Prognostic Marker in Nasopharyngeal
    Tu et al. Radiation Oncology (2015) 10:123 DOI 10.1186/s13014-015-0427-3 RESEARCH Open Access BRCC3 acts as a prognostic marker in nasopharyngeal carcinoma patients treated with radiotherapy and mediates radiation resistance in vitro Ziwei Tu1,2, Bingqing Xu1,2, Chen Qu1,2, Yalan Tao1,2, Chen Chen1,2, Wenfeng Hua2, Guokai Feng2, Hui Chang1, Zhigang Liu3, Guo Li4, Changbin Jiang5, Wei Yi5, Musheng Zeng2 and Yunfei Xia1,2* Abstract Background: BRCC3 has been found to be aberrantly expressed in breast tumors and involved in DNA damage response. The contribution of BRCC3 to nasopharyngeal carcinoma prognosis and radiosensitivity is still unclear. Methods: Immunohistochemical analysis of BRCC3 was carried out in 100 nasopharyngeal carcinoma tissues, and the protein level was correlated to patient survival. BRCC3 expression of nasopharyngeal carcinoma cell lines was determined by Western-blotting and real-time PCR. Additionally, the effects of BRCC3 knockdown on nasopharyngeal carcinoma cell clongenic survival, DNA damage repair, and cell cycle distribution after irradiation was assessed. Results: The BRCC3 protein level was inversely correlated with nasopharyngeal carcinoma patient overall survival (P < 0.001) and 3-year loco-regional relapse-free survival (P = 0.034). Multivariate analysis demonstrated that BRCC3 expression was an independent prognostic factor (P = 0.010). The expression of BRCC3 was much higher in radioresistant nasopharyngeal carcinoma cells than in radiosensitive cells. Knockdown of BRCC3 increased the cell survival fraction, attenuated DNA damage repair and resulted in G2/M cell cycle arrest in radioresistant NPC cells. Conclusions: High BRCC3 expression in nasopharyngeal carcinoma patients is associated with poor survival.
    [Show full text]
  • Literature Review of BARD1 As a Cancer Predisposing Gene with a Focus on Breast and Ovarian Cancers
    G C A T T A C G G C A T genes Review Literature Review of BARD1 as a Cancer Predisposing Gene with a Focus on Breast and Ovarian Cancers 1,2,3, 1,2, 1,2 1,2,4, Wejdan M. Alenezi y , Caitlin T. Fierheller y, Neil Recio and Patricia N. Tonin * 1 Department of Human Genetics, McGill University, Montreal, QC H3A 0G4, Canada; [email protected] (W.M.A.); caitlin.fi[email protected] (C.T.F.); [email protected] (N.R.) 2 Cancer Research Program, The Research Institute of the McGill University Health Centre, Montreal, QC H4A 3J1, Canada 3 Department of Medical Laboratory Technology, Taibah University, Medina 42353, Saudi Arabia 4 Department of Medicine, McGill University, Montreal, QC H3A 0G4, Canada * Correspondence: [email protected]; Tel.: +1-514-934-1934 (ext. 44069) Contributed equally to the work. y Received: 30 June 2020; Accepted: 23 July 2020; Published: 27 July 2020 Abstract: Soon after the discovery of BRCA1 and BRCA2 over 20 years ago, it became apparent that not all hereditary breast and/or ovarian cancer syndrome families were explained by germline variants in these cancer predisposing genes, suggesting that other such genes have yet to be discovered. BRCA1-associated ring domain (BARD1), a direct interacting partner of BRCA1, was one of the earliest candidates investigated. Sequencing analyses revealed that potentially pathogenic BARD1 variants likely conferred a low–moderate risk to hereditary breast cancer, but this association is inconsistent. Here, we review studies of BARD1 as a cancer predisposing gene and illustrate the challenge of discovering additional cancer risk genes for hereditary breast and/or ovarian cancer.
    [Show full text]
  • Fusion Protein EWS-FLI1 Is Incorporated Into a Protein Granule in Cells
    Downloaded from rnajournal.cshlp.org on September 23, 2021 - Published by Cold Spring Harbor Laboratory Press Fusion protein EWS-FLI1 is incorporated into a protein granule in cells Nasiha S. Ahmed1,2, Lucas M. Harrell2, Daniel R. Wieland2, Michelle A. Lay2, Valery F. Thompson2, Jacob C. Schwartz2* 1 Department of Molecular and Cellular Biology, The University of Arizona, Tucson, AZ 85719 2 Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ 85719 * Corresponding author: [email protected] Keywords: Ewing sarcoma, fusion proteins, phase separation, granules, transcription Nasiha S. Ahmed et al. 1 Downloaded from rnajournal.cshlp.org on September 23, 2021 - Published by Cold Spring Harbor Laboratory Press ABSTRACT Ewing sarcoma is driven by fusion proteins containing a low complexity (LC) domain that is intrinsically disordered and a powerful transcriptional regulator. The most common fusion protein found in Ewing sarcoma, EWS-FLI1, takes its LC domain from the RNA-binding protein EWSR1 (Ewing Sarcoma RNA-binding protein 1) and a DNA-binding domain from the transcription factor FLI1 (Friend Leukemia Virus Integration 1). EWS- FLI1 can bind RNA polymerase II (RNA Pol II) and self-assemble through its low-complexity (LC) domain. The ability of RNA-binding proteins like EWSR1 to self-assemble or phase separate in cells has raised questions about the contribution of this process to EWS-FLI1 activity. We examined EWSR1 and EWS-FLI1 activity in Ewing sarcoma cells by siRNA-mediated knockdown and RNA-seq analysis. More transcripts were affected by the EWSR1 knockdown than expected and these included many EWS-FLI1 regulated genes.
    [Show full text]
  • BRE Plays an Essential Role in Preventing Replicative and DNA
    www.nature.com/scientificreports OPEN BRE plays an essential role in preventing replicative and DNA damage-induced premature Received: 08 December 2015 Accepted: 08 March 2016 senescence Published: 22 March 2016 Wenting Shi1, Mei Kuen Tang1, Yao Yao1, Chengcheng Tang1, Yiu Loon Chui2 & Kenneth Ka Ho Lee1 The BRE gene, alias BRCC45, produces a 44 kDa protein that is normally distributed in both cytoplasm and nucleus. In this study, we used adult fibroblasts isolated from wild-type (WT) and BRE knockout (BRE−/−) mice to investigate the functional role of BRE in DNA repair and cellular senescence. We compared WT with BRE−/− fibroblasts at different cell passages and observed that the mutant fibroblasts entered replicative senescence earlier than the WT fibroblasts. With the use of gamma irradiation to induce DNA damage in fibroblasts, the percentage of SA-β-Gal+ cells was significantly higher in BRE−/− fibroblasts compared with WT cells, suggesting that BRE is also associated with DNA damage-induced premature senescence. We also demonstrated that the gamma irradiation induced γ-H2AX foci, a DNA damage marker, persisted significantly longer in BRE−/− fibroblasts than in WT fibroblasts, confirming that the DNA repair process is impaired in the absence of BRE. In addition, the BRCA1-A complex recruitment and homologous recombination (HR)-dependent DNA repair process upon DNA damage were impaired in BRE−/− fibroblasts. Taken together, our results demonstrate a role for BRE in both replicative senescence and DNA damage-induced premature senescence. This can be attributed to BRE being required for BRCA1-A complex-driven HR DNA repair. Cellular senescence is an irreversible physiological process that accompanies abnormal and reduced metabolic activities.
    [Show full text]
  • The Role of Bach1, Bard1 and Topbp1 Genes in Familial Breast Cancer
    D 1019 OULU 2009 D 1019 UNIVERSITY OF OULU P.O.B. 7500 FI-90014 UNIVERSITY OF OULU FINLAND ACTA UNIVERSITATIS OULUENSIS ACTA UNIVERSITATIS OULUENSIS ACTA SERIES EDITORS DMEDICA Sanna-Maria Karppinen ASCIENTIAE RERUM NATURALIUM Sanna-MariaKarppinen Professor Mikko Siponen THE ROLE OF BACH1, BARD1 BHUMANIORA AND TOPBP1 GENES IN University Lecturer Elise Kärkkäinen CTECHNICA FAMILIAL BREAST CANCER Professor Hannu Heusala DMEDICA Professor Olli Vuolteenaho ESCIENTIAE RERUM SOCIALIUM Senior Researcher Eila Estola FSCRIPTA ACADEMICA Information officer Tiina Pistokoski GOECONOMICA University Lecturer Seppo Eriksson EDITOR IN CHIEF Professor Olli Vuolteenaho PUBLICATIONS EDITOR Publications Editor Kirsti Nurkkala FACULTY OF MEDICINE, INSTITUTE OF CLINICAL MEDICINE, DEPARTMENT OF CLINICAL GENETICS, ISBN 978-951-42-9158-6 (Paperback) UNIVERSITY OF OULU; ISBN 978-951-42-9159-3 (PDF) BIOCENTER OULU, ISSN 0355-3221 (Print) UNIVERSITY OF OULU ISSN 1796-2234 (Online) ACTA UNIVERSITATIS OULUENSIS D Medica 1019 SANNA-MARIA KARPPINEN THE ROLE OF BACH1, BARD1 AND TOPBP1 GENES IN FAMILIAL BREAST CANCER Academic dissertation to be presented with the assent of the Faculty of Medicine of the University of Oulu for public defence in Auditorium 5 of Oulu University Hospital, on 26 June 2009, at 12 noon OULUN YLIOPISTO, OULU 2009 Copyright © 2009 Acta Univ. Oul. D 1019, 2009 Supervised by Professor Robert Winqvist Reviewed by Docent Outi Monni Docent Minna Tanner ISBN 978-951-42-9158-6 (Paperback) ISBN 978-951-42-9159-3 (PDF) http://herkules.oulu.fi/isbn9789514291593/ ISSN 0355-3221 (Printed) ISSN 1796-2234 (Online) http://herkules.oulu.fi/issn03553221/ Cover design Raimo Ahonen OULU UNIVERSITY PRESS OULU 2009 Karppinen, Sanna-Maria, The role of BACH1, BARD1 and TOPBP1 genes in familial breast cancer Faculty of Medicine, Institute of Clinical Medicine, Department of Clinical Genetics, University of Oulu, P.O.
    [Show full text]