DOCSLIB.ORG

Specific Synthetic Lethal Killing of RAD54B-Deficient Human Colorectal Cancer Cells by FEN1 Silencing

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Specific Synthetic Lethal Killing of RAD54B-Deficient Human Colorectal Cancer Cells by FEN1 Silencing Specific synthetic lethal killing of RAD54B-deficient human colorectal cancer cells by FEN1 silencing Kirk J. McManus, Irene J. Barrett, Yasaman Nouhi, and Philip Hieter1 Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, BC, Canada, V6T 1Z4 Communicated by Thomas D. Petes, Duke University Medical Center, Durham, NC, January 5, 2009 (received for review October 29, 2008) Mutations that cause chromosome instability (CIN) in cancer cells hypothesized that the CIN phenotype associated with tumors, produce ‘‘sublethal’’ deficiencies in an essential process (chromo- but not normal cells, represents an excellent ‘‘Achilles’ heel’’ that some segregation) and, therefore, may represent a major un- would allow for the selective killing of cancer cells. Presumably, tapped resource that could be exploited for therapeutic benefit in if cross-species tests of candidate genes can be applied to identify the treatment of cancer. If second-site unlinked genes can be second site targets that exacerbate the sublethal defect associ- identified, that when knocked down, cause a synthetic lethal (SL) ated with a CIN-inducing mutation, a novel drug target will be phenotype in combination with a somatic mutation in a CIN gene, identified. Furthermore, by generating a SL interaction network novel candidate therapeutic targets will be identified. To test this for the set of yeast CIN genes whose human homologs are idea, we took a cross species SL candidate gene approach by somatically mutated in tumors (Fig. 1B), we can identify those recapitulating a SL interaction observed between rad54 and rad27 yeast genes that are positioned as SL ‘‘interaction nodes’’ and mutations in yeast, via knockdown of the highly sequence- and whose human homologs would then represent candidate thera- functionally-related proteins RAD54B and FEN1 in a cancer cell line. peutic targets for a broad spectrum of tumors (Fig. 1 C and D). We show that knockdown of RAD54B, a gene known to be In this study, we use both RAD54B knockout and RNAi- somatically mutated in cancer, causes CIN in mammalian cells. silenced cells, and isogenic controls, to demonstrate that de- Using high-content microscopy techniques, we demonstrate that creases in human RAD54B expression in colorectal cancer cells RAD54B-deficient human colorectal cancer cells are sensitive to SL correlates with increases in chromosome numbers. RAD54B was GENETICS killing by reduced FEN1 expression, while isogenic RAD54B profi- chosen because homozygous mutations at highly conserved cient cells are not. This conserved SL interaction suggests that positions have been identified in human primary lymphoma and extrapolating SL interactions observed in model organisms for colon cancers (18), although the functional status of these homologous genes mutated in human cancers will aid in the mutant alleles has not been directly tested in a mammalian identification of novel therapeutic targets for specific killing of context (19). Furthermore, RAD54B exhibits a significant de- cancerous cells exhibiting CIN. gree of sequence and functional similarity with yeast Rdh54 and Rad54 which both exhibit strong CIN phenotypes in yeast (20). cancer therapeutics ͉ chromosome instability ͉ synthetic lethality Using a cross-species candidate gene approach and high-content digital imaging microscopy techniques, we show that the syn- enomic instability is now widely recognized as an important thetic lethality observed in yeast for rad54 rad27 double mutants Gfactor in the evolution of cancer and arises through either (see Fig. 1E and ref. 21) is conserved within a human colorectal of 2 mechanisms—increased mutation rate or chromosome Ϸ cell line (by simultaneous down-regulation of the corresponding instability (CIN). CIN correlates with 85% of solid tumors and human gene products, RAD54B and FEN1). Decreases in cell is characterized by an increased error rate in the gain or loss of numbers with concomitant increases in cellular cytotoxicity were chromosomes during cell division (1). CIN is associated with observed in RAD54B deficient/FEN1 depleted cells that were numerous different tumor types including colon (2–5), ovarian not apparent in isogenic RAD54B proficient/FEN1 depleted (6, 7), and non-Hodgkin lymphoma (8–12), and it is believed to cells. These findings represent an example of a validated target be an early event in the etiology of tumorigenesis (13–15). for selective killing of mammalian cells as a prediction from a SL Conceptually, CIN promotes tumor heterogeneity by increasing interaction between a CIN gene mutation and an unlinked gene or decreasing chromosome numbers (16), and directly affects the expression levels of both oncogenes and tumor suppressor genes mutation in yeast. We suggest that extrapolation of SL genetic encoded on the mis-segregated chromosomes. Most importantly interaction networks identified in yeast to a human context will to the work presented here, CIN gene mutations genetically provide a productive strategy to identify a broad range of novel distinguish tumor cells from normal cells and may therefore cancer therapeutic targets. represent a genetic susceptibility that could be exploited for Results selective killing (see below). Consequently, identifying the gene products that regulate chromosome stability (CS) will not only Diminished Rad54B Expression Causes Chromosome Instability in provide insights into the molecular mechanisms of chromosome Human Tissue Culture. Having previously demonstrated in yeast segregation and tumorigenesis, but it will also provide a list of that both RAD54 and RDH54 play important roles in main- candidate cancer CIN genes that may be exploited to identify taining CS (20), we wished to determine if RAD54B exhibits a similar role in humans. Accordingly, we used genomic knockouts novel therapeutic targets for the treatment of cancer. Ϫ Ϫ Ϫ In 1997, Hartwell and colleagues (17) posited that cancer cells [RAD54B / / (see SI Materials and Methods for description)] harboring somatic mutations or deletions represent genetically sensitized cells, relative to normal surrounding cells, that may be Author contributions: K.J.M. and P.H. designed research; K.J.M., I.J.B., and Y.N. performed susceptible to drug therapies selectively targeting a second research; K.J.M. contributed new reagents/analytic tools; K.J.M., I.J.B., and Y.N. analyzed unlinked gene product. They suggested that synthetic lethality data; and K.J.M. and P.H. wrote the paper. (SL), which refers to the lethal combination of 2 independently The authors declare no conflict of interest. viable mutations or deletions in 2 unlinked genes (Fig. 1A), could 1To whom correspondence should be addressed at: Michael Smith Laboratories, 2185 East be used in model organisms such as yeast to identify candidate Mall, Vancouver, BC, Canada V6T 1Z4. Email: [email protected]. SL interactions that may be conserved in humans. Because This article contains supporting information online at www.pnas.org/cgi/content/full/ chromosome segregation is an essential cellular process, we 0813414106/DCSupplemental. www.pnas.org͞cgi͞doi͞10.1073͞pnas.0813414106 PNAS Early Edition ͉ 1of6 Downloaded by guest on September 24, 2021 Fig. 1. Synthetic lethality in model organisms and human cancer. (A)ASL Fig. 2. RAD54B depletion underlies CIN. (A) Western blots depicting RAD54B interaction occurs when 2 independently viable gene mutations/deletions expression levels in knockout (RAD54Bϩ/ϩ/Ϫ and RAD54BϪ/Ϫ/Ϫ), knockdown [i.e., yfg1 (e.g., rad54) and yfg2 (e.g., rad27)] are combined to produce a lethal (RAD54B-1, RAD54B-2, and RAD54B-3), and control (Untransfected, Non- phenotype. If slow growth is observed, a synthetic growth defect (SGD) is silencing, and eGFP) cells. An ␣-tubulin loading control has been included. (B) defined. (B) A representative example of a genetic interaction network gen- DNA content analysis of RAD54B knockout and knockdown cells. Asynchro- erated from yeast data available in Biogrid (49), where circles identify genes nous cells were PI-labeled and subjected to flow cytometry. The diploid 2N and lines represent SL/SGD interactions. Note that rad27 intersects with both (G0/G1), 4N (G2/M) and Ͼ4N (aneuploid/polyploid) populations have been rad54 and rdh54. (C) Schematic representation of SL/SGD in a human cancer identified. The various cell lines/conditions are indicated in the legend. (C) context. A mutation or deletion of yfg1 (e.g., RAD54B CIN mutation) genet- Representative images of DAPI counterstained chromosomes found in mitotic ically sensitizes a cancer cell to SL attack through down-regulation of a second spreads generated from untransfected HCT116 (top left), RAD54B-1 trans- unlinked gene product [yfg2 (e.g., FEN1)], while leaving the normal adjacent fected (top right and bottom left) and RAD54BϪ/Ϫ/Ϫ (bottom right) cells. The cell(s) unaffected. (D) The yeast network presented in (B) has been humanized total chromosome numbers are indicated. (D) Scatter plots depicting the total by identifying the top hit human homolog for the respective yeast genes and chromosome distribution for cells RAD54B knockout and knockdown cells and is presented. Note that the lines only identify candidate interactions assuming controls. (E) Graphical representation of the mean chromosomes numbers evolutionary conservation. (E) Haploid rad54::URA3 and rad27::KanMX were determined for each of the conditions indicated on the x axis as quantified mated and induced to undergo meiosis. The resulting tetrads were dissected from the mitotic
Load more

Recommended publications
  • High RAD54B Expression: an Independent Predictor of Postoperative Distant Recurrence in Colorectal Cancer Patients
    www.impactjournals.com/oncotarget/ Oncotarget, Vol. 6, No. 25 High RAD54B expression: an independent predictor of postoperative distant recurrence in colorectal cancer patients Yuzo Nagai1, Yoko Yamamoto1, Takaaki Yasuhara2, Keisuke Hata1, Takeshi Nishikawa1, Toshiaki Tanaka1, Junichiro Tanaka1, Tomomichi Kiyomatsu1, Kazushige Kawai1, Hiroaki Nozawa1, Shinsuke Kazama1, Hironori Yamaguchi1, Soichiro Ishihara1, Eiji Sunami1, Takeharu Yamanaka3, Kiyoshi Miyagawa2 and Toshiaki Watanabe1 1 Department of Surgical Oncology, Faculty of Medicine, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan 2 Laboratory of Molecular Radiology, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, Japan 3 Department of Biostatistics, Graduate School of Medicine, Yokohama City University, Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, Japan Correspondence to: Toshiaki Watanabe, email: [email protected] Keywords: RAD54B, colorectal cancer, homologous recombination, prognosis, distant recurrence Received: April 21, 2015 Accepted: May 09, 2015 Published: May 20, 2015 This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. ABSTRACT We recently reported a specific mechanism that RAD54B, an important factor in homologous recombination, promotes genomic instability via the degradation of p53 protein in vitro. However, clinical significance of RAD54B in colorectal cancer (CRC) remains unclear. Thus we analyzed RAD54B gene expression in CRC patients. Using the training set (n = 123), the optimal cut-off value for stratification was determined, and validated in another cohort (n = 89). Kaplan–Meier plots showed that distant recurrence free survival was significantly lesser in high RAD54B expression group compared with that of low expression group in both training (P = 0.0013) and validation (P = 0.024) set.
    [Show full text]
  • RAD51AP1 Compensates for the Loss of RAD54 in Homology-Directed DNA Repair
    bioRxiv preprint doi: https://doi.org/10.1101/2021.07.15.452469; this version posted July 15, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. RAD51AP1 compensates for the loss of RAD54 in homology-directed DNA repair Platon Selemenakis1,2, Neelam Sharma1, Youngho Kwon3, Mollie Uhrig1, Patrick Sung3 and Claudia Wiese1* 1 Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, Colorado, 80523, USA 2 Cell and Molecular Biology Graduate Program, Colorado State University, Fort Collins, Colorado, 80523, USA 3 Department of Biochemistry and Structural Biology, University of Texas Health San Antonio, San Antonio, Texas 78229, USA * To whom correspondence should be addressed: C.W. (email: [email protected]) 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.07.15.452469; this version posted July 15, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Abstract Homology-directed repair (HDR) is a complex DNA damage repair pathway and an attractive target of inhibition in anti-cancer therapy. To develop the most efficient inhibitors of HDR in cells, it is critical to identify compensatory sub-pathways. In this study, we describe the synthetic interaction between RAD51AP1 and RAD54, two structurally unrelated proteins that function downstream of RAD51 in HDR.
    [Show full text]
  • Structure of the SWI2/SNF2 Chromatin-Remodeling Domain of Eukaryotic Rad54
    ARTICLES Structure of the SWI2/SNF2 chromatin-remodeling domain of eukaryotic Rad54 Nicolas H Thomä1, Bryan K Czyzewski1,2, Andrei A Alexeev3, Alexander V Mazin4, Stephen C Kowalczykowski3 & Nikola P Pavletich1,2 SWI2/SNF2 chromatin-remodeling proteins mediate the mobilization of nucleosomes and other DNA-associated proteins. SWI2/SNF2 proteins contain sequence motifs characteristic of SF2 helicases but do not have helicase activity. Instead, they couple ATP hydrolysis with the generation of superhelical torsion in DNA. The structure of the nucleosome-remodeling domain of zebrafish Rad54, a protein http://www.nature.com/nsmb involved in Rad51-mediated homologous recombination, reveals that the core of the SWI2/SNF2 enzymes consist of two ␣/␤-lobes similar to SF2 helicases. The Rad54 helicase lobes contain insertions that form two helical domains, one within each lobe. These insertions contain SWI2/SNF2-specific sequence motifs likely to be central to SWI2/SNF2 function. A broad cleft formed by the two lobes and flanked by the helical insertions contains residues conserved in SWI2/SNF2 proteins and motifs implicated in DNA-binding by SF2 helicases. The Rad54 structure suggests that SWI2/SNF2 proteins use a mechanism analogous to helicases to translocate on dsDNA. Cellular processes such as transcription, replication, DNA repair and breaks by Rad51-mediated homologous recombination15–20. Like other recombination require direct access to DNA. This process is facilitated SWI2/SNF2 remodeling enzymes, Rad54 can translocate on DNA, gen- by the SWI2/SNF2 family of ATPases, which detach DNA from histones erate superhelical torsion and enhance the accessibility to nucleosomal and other bound proteins1,2. The SWI2/SNF2 chromatin remodeling DNA18,19,21.
    [Show full text]
  • Werner Syndrome Protein Participates in a Complex with RAD51, RAD54
    Erratum Werner syndrome protein participates in a complex with RAD51, RAD54, RAD54B and ATR in response to ICL-induced replication arrest Marit Otterlei, Per Bruheim, Byungchan Ahn, Wendy Bussen, Parimal Karmakar, Kathy Baynton and Vilhelm A. Bohr Journal of Cell Science 119, 5215 (2006) doi:10.1242/jcs.03359 There was an error in the first e-press version of the article published in J. Cell Sci. 119, 5137-5146. The first e-press version of this article gave the page range as 5114-5123, whereas it should have been 5137-5146. We apologise for this mistake. Research Article 5137 Werner syndrome protein participates in a complex with RAD51, RAD54, RAD54B and ATR in response to ICL-induced replication arrest Marit Otterlei1,2,*, Per Bruheim1,3,§, Byungchan Ahn1,4,§, Wendy Bussen5, Parimal Karmakar1,6, Kathy Baynton7 and Vilhelm A. Bohr1 1Laboratory of Molecular Gerontology, National Institute on Aging, NIH, 5600 Nathan Shock Dr., Baltimore, MD 21224, USA 2Department of Cancer Research and Molecular Medicine, Laboratory Centre, Faculty of Medicine, Norwegian University of Science and Technology, Erling Skjalgsons gt. 1, N-7006 Trondheim, Norway 3Department of Biotechnology, Norwegian University of Science and Technology, N-7491 Trondheim, Norway 4Department of Life Sciences, University of Ulsan, Ulsan 680-749, Korea 5Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, 333 Cedar St, SHM-C130, New Haven, CT 06515, USA 6Department of Life Science and Biotechnology, Jadavpur University, Kolkata-700 032, WB,
    [Show full text]
  • Role of Rad54, Rad54b and Snm1 in DNA Damage Repair
    Role of Rad54, Rad54B and Snm1 in DNA Damage Repair De rol van Rad54, Rad54B en Snm1 in herstel van schade aan DNA Proefschrift Ter verkrijging van de graad van doctor aan de Erasmus Universiteit Rotterdam op gezag van de Rector Magnificus Prof.dr.ir. J.H. van Bemmel en volgens besluit van het College voor Promoties. De openbare verdediging zal plaatsvinden op woensdag 8 oktober 2003 om 9.45 uur door Joanna Wesoty Geboren te Poznan, Polen Promotiecommissie Promotoren: Prof.dr. R. Kanaar Prof.dr. J.H.J. Hoeijmakers Overige !eden: Prof.dr. J.A. Grootegoed Dr. G.T.J. van der Horst Prof.dr.ir. A.A. van Zeeland The work presented in this thesis was performed at the Department of Cell Biology and Genetics of Erasmus MC in Rotterdam from 1998 to 2003. The research and the printing of the thesis were financially supported by the Dutch Cancer Society. Dziadkowi i Jasiowi Contents list of abbreviations 6 Scope of the thesis 7 Chapter 1 11 DNA double-strand breaks: significance, generation, repair and link to cellular processes 1. Generation of DSBs, their significance and repair 11 2. Homologous recombination and non-homologous end joining - interplay and contribution to repair of DBSs 13 3. Biochemistry of homologous recombination 15 4. DSBs in meiosis 23 5. Non-homologous end joining 26 6. DNA rearrangements in the immune system 27 7. lnterstrand cross-links and homologous recombination 33 8. Perspective 34 Chapter 2 37 Analysis of mouse Rad54 expression and its implications for homologous recombination Chapter 3 57 Sublethal damage recovery
    [Show full text]
  • The RAD51-Stimulatory Compound RS-1 Can Exploit the RAD51 Overexpression That Exists in Cancer Cells and Tumors
    Author Manuscript Published OnlineFirst on April 21, 2014; DOI: 10.1158/0008-5472.CAN-13-3220 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited. The RAD51-stimulatory compound RS-1 can exploit the RAD51 overexpression that exists in cancer cells and tumors Jennifer M Mason1,2, Hillary L. Logan1,2, Brian Budke1, Megan Wu1, Michal Pawlowski3, Ralph R. Weichselbaum1,4, Alan P. Kozikowski3, Douglas K. Bishop1,5,6, and Philip P. Connell1,6 1Department of Radiation and Cellular Oncology, 4Ludwig Center for Metastasis Research, 5Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL 3Department of Medicinal Chemistry and Pharmacognosy, Drug Discovery Program, University of Illinois at Chicago, Chicago, IL 2These authors contributed equally 6These authors contributed equally Precis: We developed a novel therapeutic strategy that exploits the high levels of RAD51 overexpression that occur in human cancers, using the RAD51-stimulatory compound RS-1 to form genotoxic RAD51 protein complexes on undamaged chromatin. Key Words: DNA repair, Homologous recombination, RAD51, RS-1 Conflicts of interest: The authors disclose no potential conflicts of interest Running Title: RAD51-stimulatory compound kills malignant cells Correspondence should be directed to: Philip P. Connell MD Dep. of Radiation and Cellular Oncology University of Chicago 5758 S. Maryland Ave, MC 9006 Chicago, IL 60637 Phone: (773) 834-8119 Fax: (773) 702-0610 email: [email protected] 1 Downloaded from cancerres.aacrjournals.org on October 2, 2021. © 2014 American Association for Cancer Research. Author Manuscript Published OnlineFirst on April 21, 2014; DOI: 10.1158/0008-5472.CAN-13-3220 Author manuscripts have been peer reviewed and accepted for publication but have not yet been edited.
    [Show full text]
  • DNA Repair Gene Knockdown Cell Lines Tools to Study Genomic Instability and Genotoxic Stress DNA Repair Proteins Maintain the Stability of the Genome
    FORM TCA 5 DATE Trevigen Cell Assays (TCA), a division of Trevigen, Inc., was established in 2008 to conduct contract research for the pharmaceutical, biotechnology, government and academic segments of the medical research market. TCA specializes in designing and conducting assays for lead compounds and genotoxic screening based on DNA damage and repair as well as cancer cell behavior. DNA Repair Gene Knockdown Cell Lines Tools to study genomic instability and genotoxic stress DNA repair proteins maintain the stability of the genome. When repair protein function is impaired through mutations, the genome may become unstable which is a hallmark of solid tumors. The availability of this panel of knockdown cell lines will permit scientists to study the molecular etiology of tumor genomic instability and to exploit it in oncology research. The Knockdown line encompasses the Base Excision Repair pathway, Non-Homologous End Joining, Mismatch Repair and Homologous Recombination pathways. Base Excision Repair - PARP1, OGG1, NTHL1, NEIL2, NEIL3, UNG, SMUG1, MPG, MutYH, Neil1, PARP2, PARP3, XRCC1, APE1, APE2, TDG, MBD4, LIG3 Mismatch Repair - MSH2, PMS2, MLH3, MSH3, MLH1, MSH5, MSH6 Non-homologous End Joining - SIRT5, SIRT1, PRKDC, SIRT2, XRCC6 (Ku70), XRCC5 (Ku80), SIRT3 Homologous Recombination - RAD54B, NBS1, RAD21, XRCC3, SMC6L1, SHFM1, BRCA1, RAD51C, RAD50 Trevigen’s Knockdown Cell Lines are target specific LN428 (glioblastoma) shRNA lentivirus transduced cells. They are rigorously qualified and mycoplasma free. The percent knockdown levels range from 63-98% depending on the gene, as evaluated by RT-PCR. Lentiviruses are maintained by puromycin selection. Features • Tested negative for Mycoplasma • Easy to use Glioblastoma based knock down cells with reliable short and long term knockdown efficiency.
    [Show full text]
  • RAD51AP1 and RAD54 Underpin Two Distinct RAD51-Dependent Routes of DNA Damage Repair Via Homologous Recombination
    bioRxiv preprint doi: https://doi.org/10.1101/2021.07.15.452469; this version posted August 17, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. RAD51AP1 and RAD54 underpin two distinct RAD51-dependent routes of DNA damage repair via homologous recombination Platon Selemenakis1,2,$, Neelam Sharma1, Youngho Kwon3, Mollie Uhrig1, Patrick Sung3 and Claudia Wiese1* 1 Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, Colorado 80523, USA 2 Cell and Molecular Biology Graduate Program, Colorado State University, Fort Collins, Colorado 80523, USA 3 Department of Biochemistry and Structural Biology, University of Texas Health San Antonio, San Antonio, Texas 78229, USA $ Present address: Department of Cancer Biology, University of Texas MD Anderson Cancer Center, Houston, Texas 77054, USA * To whom correspondence should be addressed: C.W. (email: [email protected]) 1 bioRxiv preprint doi: https://doi.org/10.1101/2021.07.15.452469; this version posted August 17, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. Abstract Homologous recombination (HR) is a complex DNA damage repair pathway and an attractive target of inhibition in anti-cancer therapy. To help guide the development of efficient HR inhibitors, it is critical to identify compensatory sub-pathways.
    [Show full text]
  • Identification of RAD54 Homolog B As a Promising Therapeutic Target for Breast Cancer
    5350 ONCOLOGY LETTERS 18: 5350-5362, 2019 Identification of RAD54 homolog B as a promising therapeutic target for breast cancer JING FENG1,2*, JUANJUAN HU1,2* and YING XIA1,2 1Institute of Chemical Component Analysis of Traditional Chinese Medicine, Chongqing Medical and Pharmaceutical College; 2Engineering Research Center of Pharmaceutical Sciences, Chongqing 401331, P.R. China Received January 24, 2019; Accepted July 26, 2019 DOI: 10.3892/ol.2019.10854 Abstract. Breast cancer is a recognized threat to the health Introduction of women globally. Due to the lack of the knowledge about the molecular pathogenesis of breast cancer, therapeutic Cancer is considered to be one of the most dangerous factors strategies remain inadequate, especially for aggressive breast to human life. The global cancer statistics for 2018 demon- cancer. In the present study, sequential bioinformatics analysis strated that breast cancer exhibits the highest morbidity and was performed using data from the GSE20711 dataset, and mortality rates in females worldwide compared with other the results demonstrated that three genes may impact the types of cancer (1). Several therapeutic strategies have been survival of patients with breast cancer. One of these genes, developed for breast cancer treatment, including surgery, RAD54 homolog B (RAD54B), may be a potential prognostic chemotherapy, radiotherapy, hormone therapy and newly factor for breast cancer. A signature was established that improved immunotherapy (2). However, due to the high could evaluate the overall survival for patients with breast heterogeneity among different types of breast cancer, the cancer based on the risk score calculated from RAD54B prognosis for a number of patients is still poor, especially for expression and the Tumor-Node-Metastasis (TNM) stage patients with distant metastases, who are usually diagnosed [risk score=expRAD54B x 0.236 + TNM stage (I/II=0 or at a late stage (3).
    [Show full text]
  • PARP1 Bound to XRCC2 Promotes Tumor Progression in Colorectal Cancer
    PARP1 Bound to XRCC2 Promotes Tumor Progression in Colorectal Cancer Kaiwu Xu Hunan Provincial People's Hospital Zhige Yu Hunan Provincial People's Hospital Tailiang Lu Hunan Provincial People's Hospital Wei Peng Hunan Provincial People's Hospital Yongqiang Gong Hunan Provincial People's Hospital Chaowu Chen ( [email protected] ) Hunan Provincial People's Hospital https://orcid.org/0000-0001-5912-9050 Research Keywords: PARP1, XRCC2, Progression, Colorectal cancer, Gastrointestinal Posted Date: February 4th, 2021 DOI: https://doi.org/10.21203/rs.3.rs-176757/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Page 1/18 Abstract Background: By complexing poly (ADP-ribose) (PAR) in reaction to breaked strand, PAR polymerase1 (PARP1) acts as the key enzyme participated in DNA repair. However, recent studies suggest that unrepaired DNA breaks results in persistent PARP1 activation, which leads to a progressively reduce in hexokinase1 (HK1) activity and cell death. So the molecular mechanism of PARP1 remains elusive. Methods: 212 colorectal cancer (CRC) patients who had the operation at our hospital were recruited. Used immunohistochemistry to evaluate PARP1 expression. Survival analysis was calculated based on PARP1 expression. Results: Compared with matching adjacent noncancerous tissue, in CRC tissue, PARP1 expression was remarkably higher, which was correlated with the degree of differentiation, TNM stage, depth of invasion, distant metastasis, and 5-year survival. Furthermore, after constructing CRC cell lines stably expressing low or high PARP1, we found that PARP1 overexpression promoted proliferation, and proved PARP1 interacted with XRCC2 in CRC cells by immunoprecipitation (IP) analysis.
    [Show full text]
  • Are Pathogenic Germline Variants in Metastatic Melanoma Associated with Resistance to Combined Immunotherapy?
    cancers Article Are Pathogenic Germline Variants in Metastatic Melanoma Associated with Resistance to Combined Immunotherapy? Teresa Amaral 1,2 , Martin Schulze 3, Tobias Sinnberg 1 , Maike Nieser 3, Peter Martus 4 , Florian Battke 5 , Claus Garbe 1, Saskia Biskup 3,5 and Andrea Forschner 1,* 1 Center for Dermatooncology, Department of Dermatology, University Hospital Tuebingen, Eberhard Karls University, 72076 Tuebingen, Germany; [email protected] (T.A.); [email protected] (T.S.); [email protected] (C.G.) 2 Portuguese Air Force, Health Care Direction, 1649-020 Lisbon, Portugal 3 Practice for Human Genetics, 72076 Tuebingen, Germany; [email protected] (M.S.); [email protected] (M.N.); [email protected] (S.B.) 4 Institute for Clinical Epidemiology and applied Biostatistics (IKEaB), Eberhard Karls University, 72076 Tuebingen, Germany; [email protected] 5 Center for Genomics and Transcriptomics (CeGaT) GmbH, 72076 Tuebingen, Germany; [email protected] * Correspondence: [email protected]; Tel.: +49-(0)-7071-29 84555; Fax: +49-(0)-7071-29-4599 Received: 24 March 2020; Accepted: 27 April 2020; Published: 28 April 2020 Abstract: Background: Combined immunotherapy has significantly improved survival of patients with advanced melanoma, but there are still patients that do not benefit from it. Early biomarkers that indicate potential resistance would be highly relevant for these patients. Methods: We comprehensively analyzed tumor and blood samples from patients with advanced melanoma, treated with combined immunotherapy and performed descriptive and survival analysis. Results: Fifty-nine patients with a median follow-up of 13 months (inter quartile range (IQR) 11–15) were included.
    [Show full text]
  • Specific Synthetic Lethal Killing of RAD54B-Deficient Human Colorectal Cancer Cells by FEN1 Silencing
    Specific synthetic lethal killing of RAD54B-deficient human colorectal cancer cells by FEN1 silencing Kirk J. McManus, Irene J. Barrett, Yasaman Nouhi, and Philip Hieter1 Michael Smith Laboratories, University of British Columbia, 2185 East Mall, Vancouver, BC, Canada, V6T 1Z4 Communicated by Thomas D. Petes, Duke University Medical Center, Durham, NC, January 5, 2009 (received for review October 29, 2008) Mutations that cause chromosome instability (CIN) in cancer cells hypothesized that the CIN phenotype associated with tumors, produce ‘‘sublethal’’ deficiencies in an essential process (chromo- but not normal cells, represents an excellent ‘‘Achilles’ heel’’ that some segregation) and, therefore, may represent a major un- would allow for the selective killing of cancer cells. Presumably, tapped resource that could be exploited for therapeutic benefit in if cross-species tests of candidate genes can be applied to identify the treatment of cancer. If second-site unlinked genes can be second site targets that exacerbate the sublethal defect associ- identified, that when knocked down, cause a synthetic lethal (SL) ated with a CIN-inducing mutation, a novel drug target will be phenotype in combination with a somatic mutation in a CIN gene, identified. Furthermore, by generating a SL interaction network novel candidate therapeutic targets will be identified. To test this for the set of yeast CIN genes whose human homologs are idea, we took a cross species SL candidate gene approach by somatically mutated in tumors (Fig. 1B), we can identify those recapitulating a SL interaction observed between rad54 and rad27 yeast genes that are positioned as SL ‘‘interaction nodes’’ and mutations in yeast, via knockdown of the highly sequence- and whose human homologs would then represent candidate thera- functionally-related proteins RAD54B and FEN1 in a cancer cell line.
    [Show full text]