TY - JOUR
T1 - The Long Noncoding RNA CCAT2 Induces Chromosomal Instability Through BOP1-AURKB Signaling
AU - Chen, Baoqing
AU - Dragomir, Mihnea P.
AU - Fabris, Linda
AU - Bayraktar, Recep
AU - Knutsen, Erik
AU - Liu, Xu
AU - Tang, Changyan
AU - Li, Yongfeng
AU - Shimura, Tadanobu
AU - Ivkovic, Tina Catela
AU - Cruz De los Santos, Mireia
AU - Anfossi, Simone
AU - Shimizu, Masayoshi
AU - Shah, Maitri Y.
AU - Ling, Hui
AU - Shen, Peng
AU - Multani, Asha S.
AU - Pardini, Barbara
AU - Burks, Jared K.
AU - Katayama, Hiroyuki
AU - Reineke, Lucas C.
AU - Huo, Longfei
AU - Syed, Muddassir
AU - Song, Shumei
AU - Ferracin, Manuela
AU - Oki, Eiji
AU - Fromm, Bastian
AU - Ivan, Cristina
AU - Bhuvaneshwar, Krithika
AU - Gusev, Yuriy
AU - Mimori, Koshi
AU - Menter, David
AU - Sen, Subrata
AU - Matsuyama, Takatoshi
AU - Uetake, Hiroyuki
AU - Vasilescu, Catalin
AU - Kopetz, Scott
AU - Parker-Thornburg, Jan
AU - Taguchi, Ayumu
AU - Hanash, Samir M.
AU - Girnita, Leonard
AU - Slaby, Ondrej
AU - Goel, Ajay
AU - Varani, Gabriele
AU - Gagea, Mihai
AU - Li, Chunlai
AU - Ajani, Jaffer A.
AU - Calin, George A.
N1 - Funding Information:
Funding Dr Calin is the Felix L. Haas Endowed Professor in Basic Science. Work in Dr Calin’s laboratory is supported by National Institutes of Health (NIH), National Center for Advancing Translational Sciences grant UH3TR00943-01 through the NIH Common Fund, Office of Strategic Coordination , National Cancer Institute (NCI) grants 1R01 CA182905-01 and 1R01CA222007-01A1 , a National Institute of General Medical Sciences (NIGMS) 1R01GM122775-01 grant, a U54 grant #CA096297 / CA096300 –University of Puerto Rico and The University of Texas MD Anderson Cancer Partnership for Excellence in Cancer Research 2016 Pilot Project, a Team DOD (CA160445P1) grant, a Chronic Lymphocytic Leukemia Moonshot Flagship project, the UT MD Anderson Cancer Center Duncan Family Institute for Cancer Prevention and Risk Assessment, a Sister Institution Network Fund 2017 grant, and the Estate of C. G. Johnson Jr. Dr Pardini is recipient of a Fulbright Research Scholarship (2018). Work at the University of Washington was supported by NIGMS R35 GM121487 . The work of Dr Baoqing Chen is supported by National Natural Science Foundation of China (no. 81902462 ). The work in Dr. Goel’s laboratory is supported by the grants CA72851 , CA181572 , and CA202797 from the NCI and NIH . The work of Dr Parker-Thornburg was supported by the grants 5R50CA211121-03 and Cancer Center Support Grant for the genetically engineered mouse facility core.
Publisher Copyright:
© 2020 AGA Institute
PY - 2020/12
Y1 - 2020/12
N2 - 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.
AB - 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.
KW - Aneuploidy
KW - MSS
KW - Noncoding RNA
KW - Tumorigenesis
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U2 - 10.1053/j.gastro.2020.08.018
DO - 10.1053/j.gastro.2020.08.018
M3 - Article
C2 - 32805281
AN - SCOPUS:85092523174
SN - 0016-5085
VL - 159
SP - 2146-2162.e33
JO - Gastroenterology
JF - Gastroenterology
IS - 6
ER -