BACKGROUND & AIMS: Chromosomal instability (CIN) is a carcinogenesis event that promotes metastasis and resistance to therapy by unclear mechanisms. Expression of the colon cancer-associated transcript 2 gene (CCAT2), which encodes a long noncoding RNA (lncRNA), associates with CIN, but little is known about how CCAT2 lncRNA regulates this cancer enabling characteristic. METHODS: We performed cytogenetic analysis of colorectal cancer (CRC) cell lines (HCT116, KM12C/SM, and HT29) overexpressing CCAT2 and colon organoids from C57BL/6N mice with the CCAT2 transgene and without (controls). CRC cells were also analyzed by immunofluorescence microscopy, γ-H2AX, and senescence assays. CCAT2 transgene and control mice were given azoxymethane and dextran sulfate sodium to induce colon tumors. We performed gene expression array and mass spectrometry to detect downstream targets of CCAT2 lncRNA. We characterized interactions between CCAT2 with downstream proteins using MS2 pull-down, RNA immunoprecipitation, and selective 2'-hydroxyl acylation analyzed by primer extension analyses. Downstream proteins were overexpressed in CRC cells and analyzed for CIN. Gene expression levels were measured in CRC and non-tumor tissues from 5 cohorts, comprising more than 900 patients. RESULTS: High expression of CCAT2 induced CIN in CRC cell lines and increased resistance to 5-fluorouracil and oxaliplatin. Mice that expressed the CCAT2 transgene developed chromosome abnormalities, and colon organoids derived from crypt cells of these mice had a higher percentage of chromosome abnormalities compared with organoids from control mice. The transgenic mice given azoxymethane and dextran sulfate sodium developed more and larger colon polyps than control mice given these agents. Microarray analysis and mass spectrometry indicated that expression of CCAT2 increased expression of genes involved in ribosome biogenesis and protein synthesis. CCAT2 lncRNA interacted directly with and stabilized BOP1 ribosomal biogenesis factor (BOP1). CCAT2 also increased expression of MYC, which activated expression of BOP1. Overexpression of BOP1 in CRC cell lines resulted in chromosomal missegregation errors, and increased colony formation, and invasiveness, whereas BOP1 knockdown reduced viability. BOP1 promoted CIN by increasing the active form of aurora kinase B, which regulates chromosomal segregation. BOP1 was overexpressed in polyp tissues from CCAT2 transgenic mice compared with healthy tissue. CCAT2 lncRNA and BOP1 mRNA or protein were all increased in microsatellite stable tumors (characterized by CIN), but not in tumors with microsatellite instability compared with nontumor tissues. Increased levels of CCAT2 lncRNA and BOP1 mRNA correlated with each other and with shorter survival times of patients. CONCLUSIONS: We found that overexpression of CCAT2 in colon cells promotes CIN and carcinogenesis by stabilizing and inducing expression of BOP1 an activator of aurora kinase B. Strategies to target this pathway might be developed for treatment of patients with microsatellite stable colorectal tumors.

Center for Gastrointestinal Research; Center for Translational Genomics and Oncology Baylor Scott and White Research Institute Charles A Sammons Cancer Center Baylor University Medical Center Dallas Texas

Central European Institute of Technology Masaryk University Brno Czech Republic

Central European Institute of Technology Masaryk University Brno Czech Republic; Department of Biology Faculty of Medicine Masaryk University Brno Czech Republic

Department of Chemistry University of Washington Seattle Washington

Department of Clinical Cancer Prevention University of Texas MD Anderson Cancer Center Houston Texas

Department of Experimental Diagnostic and Specialty Medicine University of Bologna Bologna Italy

Department of Experimental Therapeutics The University of Texas MD Anderson Cancer Center Houston Texas

Department of Experimental Therapeutics The University of Texas MD Anderson Cancer Center Houston Texas; Center for RNA Interference and Non Coding RNAs The University of Texas MD Anderson Cancer Center Houston Texas

Department of Experimental Therapeutics The University of Texas MD Anderson Cancer Center Houston Texas; Department of General Surgery Fundeni Clinical Hospital Carol Davila University of Medicine and Pharmacy Bucharest Romania

Department of Experimental Therapeutics The University of Texas MD Anderson Cancer Center Houston Texas; Department of Medical Biology Faculty of Health Sciences The Arctic University of Norway Tromsø Norway

Department of Experimental Therapeutics The University of Texas MD Anderson Cancer Center Houston Texas; Department of Oncology Nanfang Hospital Southern Medical University Guangzhou China

Department of Experimental Therapeutics The University of Texas MD Anderson Cancer Center Houston Texas; Department of Radiation Oncology State Key Laboratory of Oncology in South China Collaborative Innovation Center for Cancer Medicine Sun Yat sen University Cancer Center Guangzhou Guangdong China; Department of Thoracic Oncology Cancer Center and State Key Laboratory of Biotherapy West China Hospital Sichuan University Chengdu Sichuan China

Department of Experimental Therapeutics The University of Texas MD Anderson Cancer Center Houston Texas; Department of Thoracic Surgery The 1st Affiliated Hospital of Xi'an Jiaotong University Xi'an China

Department of Gastrointestinal Medical Oncology The University of Texas MD Anderson Cancer Center Houston Texas

Department of Gastrointestinal Surgery Tokyo Medical and Dental University Graduate School of Medicine Tokyo Japan

Department of General Surgery Fundeni Clinical Hospital Carol Davila University of Medicine and Pharmacy Bucharest Romania; Carol Davila University of Medicine and Pharmacy Bucharest Romania

Department of Genetics The University of Texas MD Anderson Cancer Center Houston Texas

Department of Leukemia The University of Texas MD Anderson Cancer Center Houston Texas

Department of Neuroscience Baylor College of Medicine Houston Texas

Department of Oncology Pathology Bioclinicum Karolinska Institute and Karolinska University Hospital Stockholm Sweden

Department of Specialized Surgeries Graduate School Tokyo Medical and Dental University Tokyo Japan

Department of Surgery and Science Graduate School of Medical Sciences Kyushu University Fukuoka Japan

Department of Surgery Kyushu University Beppu Hospital Beppu Japan

Department of Translational Molecular Pathology The University of Texas MD Anderson Cancer Center Houston Texas

Department of Veterinary Medicine and Surgery The University of Texas MD Anderson Cancer Center Houston Texas

Innovation Center for Biomedical Informatics Georgetown University Washington District of Columbia

Science for Life Laboratory Department of Molecular Biosciences The Wenner Gren Institute Stockholm University Stockholm Sweden; Department of Tumor Biology Institute for Cancer Research The Norwegian Radium Hospital Oslo University Hospital Oslo Norway

Zobrazit více v PubMed

Gordon DJ, Resio B, Pellman D. Causes and consequences of aneuploidy in cancer. Nat Rev Genet 2012;13:189–203. PubMed

Boveri M. Über Mitosen bei einseitiger Chromosomenbindung. Jenaische Zeitschrift für Naturwissenschaft 1903;37:401–443.

Negrini S, Gorgoulis VG, Halazonetis TD. Genomic instability--an evolving hallmark of cancer. Nat Rev Mol Cell Biol 2010;11:220–8. PubMed

Gronroos E, Lopez-Garcia C. Tolerance of Chromosomal Instability in Cancer: Mechanisms and Therapeutic Opportunities. Cancer Res 2018;78:6529–6535. PubMed

Galimberti F, Thompson SL, Ravi S, et al. Anaphase catastrophe is a target for cancer therapy. Clin Cancer Res 2011;17:1218–22. PubMed PMC

Hoffelder DR, Luo L, Burke NA, et al. Resolution of anaphase bridges in cancer cells. Chromosoma 2004;112:389–97. PubMed

Bakhoum SF, Cantley LC. The Multifaceted Role of Chromosomal Instability in Cancer and Its Microenvironment. Cell 2018;174:1347–1360. PubMed PMC

Dragomir MP, Kopetz S, Ajani JA, et al. Non-coding RNAs in GI cancers: from cancer hallmarks to clinical utility. Gut 2020;69:748–763. PubMed

Ling H, Spizzo R, Atlasi Y, et al. CCAT2, a novel noncoding RNA mapping to 8q24, underlies metastatic progression and chromosomal instability in colon cancer. Genome Res 2013;23:1446–61. PubMed PMC

Redis RS, Vela LE, Lu W, et al. Allele-Specific Reprogramming of Cancer Metabolism by the Long Non-coding RNA CCAT2. Mol Cell 2016;61:520–534. PubMed PMC

Shah MY, Ferracin M, Pileczki V, et al. Cancer-associated rs6983267 SNP and its accompanying long noncoding RNA CCAT2 induce myeloid malignancies via unique SNP-specific RNA mutations. Genome Res 2018;28:432–447. PubMed PMC

Pellagatti A, Boultwood J. The molecular pathogenesis of the myelodysplastic syndromes. Eur J Haematol 2015;95:3–15. PubMed

Maslov AY, Vijg J. Genome instability, cancer and aging. Biochim Biophys Acta 2009;1790:963–9. PubMed PMC

Bayani J, Selvarajah S, Maire G, et al. Genomic mechanisms and measurement of structural and numerical instability in cancer cells. Seminars in Cancer Biology 2007;17:5–18. PubMed

Gerling M, Glauben R, Habermann JK, et al. Characterization of chromosomal instability in murine colitis-associated colorectal cancer. PLoS One 2011;6:e22114. PubMed PMC

Roux PP, Topisirovic I. Regulation of mRNA translation by signaling pathways. Cold Spring Harb Perspect Biol 2012;4. PubMed PMC

Kim TH, Leslie P, Zhang Y. Ribosomal proteins as unrevealed caretakers for cellular stress and genomic instability. Oncotarget 2014;5:860–71. PubMed PMC

Killian A, Sarafan-Vasseur N, Sesboue R, et al. Contribution of the BOP1 gene, located on 8q24, to colorectal tumorigenesis. Genes Chromosomes Cancer 2006;45:874–81. PubMed

Killian A, Le Meur N, Sesboue R, et al. Inactivation of the RRB1-Pescadillo pathway involved in ribosome biogenesis induces chromosomal instability. Oncogene 2004;23:8597–602. PubMed

Beroukhim R, Mermel CH, Porter D, et al. The landscape of somatic copy-number alteration across human cancers. Nature 2010;463:899–905. PubMed PMC

Rohrmoser M, Holzel M, Grimm T, et al. Interdependence of Pes1, Bop1, and WDR12 controls nucleolar localization and assembly of the PeBoW complex required for maturation of the 60S ribosomal subunit. Mol Cell Biol 2007;27:3682–94. PubMed PMC

Neer EJ, Schmidt CJ, Nambudripad R, et al. The ancient regulatory-protein family of WD-repeat proteins. Nature 1994;371:297–300. PubMed

Kent WJ, Sugnet CW, Furey TS, et al. The human genome browser at UCSC. Genome Res 2002;12:996–1006. PubMed PMC

Qi J, Yu Y, Akilli Ozturk O, et al. New Wnt/beta-catenin target genes promote experimental metastasis and migration of colorectal cancer cells through different signals. Gut 2016;65:1690–701. PubMed

Grummt I. The nucleolus-guardian of cellular homeostasis and genome integrity. Chromosoma 2013;122:487–97. PubMed

Song L, Bhuvaneshwar K, Wang Y, et al. CINdex: A Bioconductor Package for Analysis of Chromosome Instability in DNA Copy Number Data. Cancer Inform 2017;16:1176935117746637. PubMed PMC

Yasui Y, Urano T, Kawajiri A, et al. Autophosphorylation of a newly identified site of Aurora-B is indispensable for cytokinesis. J Biol Chem 2004;279:12997–3003. PubMed

den Hollander J, Rimpi S, Doherty JR, et al. Aurora kinases A and B are up-regulated by Myc and are essential for maintenance of the malignant state. Blood 2010;116:1498–505. PubMed PMC

Jambhekar A, Emerman AB, Schweidenback CT, et al. RNA stimulates Aurora B kinase activity during mitosis. PLoS One 2014;9:e100748. PubMed PMC

Cancer Genome Atlas Research N. Comprehensive molecular characterization of gastric adenocarcinoma. Nature 2014;513:202–9. PubMed PMC

DeRose YS, Wang G, Lin YC, et al. Tumor grafts derived from women with breast cancer authentically reflect tumor pathology, growth, metastasis and disease outcomes. Nat Med 2011;17:1514–20. PubMed PMC

Simons CC, Hughes LA, Smits KM, et al. A novel classification of colorectal tumors based on microsatellite instability, the CpG island methylator phenotype and chromosomal instability: implications for prognosis. Ann Oncol 2013;24:2048–56. PubMed

Watanabe T, Kobunai T, Yamamoto Y, et al. Chromosomal instability (CIN) phenotype, CIN high or CIN low, predicts survival for colorectal cancer. J Clin Oncol 2012;30:2256–64. PubMed

Jamal-Hanjani M, Wilson GA, McGranahan N, et al. Tracking the Evolution of Non-Small-Cell Lung Cancer. N Engl J Med 2017;376:2109–2121. PubMed

Carter SL, Eklund AC, Kohane IS, et al. A signature of chromosomal instability inferred from gene expression profiles predicts clinical outcome in multiple human cancers. Nature Genetics 2006;38:1043–1048. PubMed

Lips EH, van Eijk R, de Graaf EJ, et al. Integrating chromosomal aberrations and gene expression profiles to dissect rectal tumorigenesis. BMC Cancer 2008;8:314. PubMed PMC

Carmena M, Wheelock M, Funabiki H, et al. The chromosomal passenger complex (CPC): from easy rider to the godfather of mitosis. Nat Rev Mol Cell Biol 2012;13:789–803. PubMed PMC

Munoz-Barrera M, Monje-Casas F. Increased Aurora B activity causes continuous disruption of kinetochore-microtubule attachments and spindle instability. Proc Natl Acad Sci U S A 2014;111:E3996–4005. PubMed PMC