The Long Noncoding RNA CCAT2 Induces Chromosomal Instability Through BOP1-AURKB Signaling.
The Long Noncoding RNA CCAT2 Induces Chromosomal Instability Through BOP1-AURKB Signaling.
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DOI：
10.1053/j.gastro.2020.08.018
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发表时间：
2020-12
期刊：
影响因子：
29.4
通讯作者：
Calin GA
中科院分区：
文献类型：
--
作者：
Chen B;Dragomir MP;Fabris L;Bayraktar R;Knutsen E;Liu X;Tang C;Li Y;Shimura T;Ivkovic TC;De Los Santos MC;Anfossi S;Shimizu M;Shah MY;Ling H;Shen P;Multani AS;Pardini B;Burks JK;Katayama H;Reineke LC;Huo L;Syed M;Song S;Ferracin M;Oki E;Fromm B;Ivan C;Bhuvaneshwar K;Gusev Y;Mimori K;Menter D;Sen S;Matsuyama T;Uetake H;Vasilescu C;Kopetz S;Parker-Thornburg J;Taguchi A;Hanash SM;Girnita L;Slaby O;Goel A;Varani G;Gagea M;Li C;Ajani JA;Calin GA
染色体不稳定(CIN)是一种致癌事件，可促进肿瘤转移和抵抗治疗，其机制尚不清楚。结肠癌相关转录本2基因(CCAT2)编码一个长的非编码RNA(LncRNA)，它的表达与CIN有关，但对CCAT2 lncRNA如何调节这一致癌特性知之甚少。我们对高表达CCAT2的结直肠癌(CRC)细胞系(HCT116、KM12C/SM和HT29)和来自C57BL/6N小鼠(对照组)的CCAT2转基因小鼠和结肠类器官进行了细胞遗传学分析。用免疫荧光显微镜、γ-H_2AX和衰老实验对结直肠癌细胞进行分析。用偶氮甲烷和葡聚糖硫酸钠(DSS)诱导CCAT2转基因小鼠和对照组小鼠结肠癌。我们进行了基因表达阵列和质谱学检测CCAT2 lncRNA下游靶点。我们用MS2下拉、RNA免疫沉淀和SHAPE分析表征了CCAT2与下游蛋白质之间的相互作用。下游蛋白在结直肠癌细胞中过表达，并分析CIN。对来自5个队列的超过900名患者的结直肠癌和非肿瘤组织的基因表达水平进行了测量。CCAT2高表达可诱导结直肠癌细胞发生CIN，并增加对5-氟尿嘧啶和奥沙利铂的耐药性。表达CCAT2转基因的小鼠出现了染色体异常，这些小鼠的隐窝细胞来源的结肠器官与对照组小鼠的器官相比，染色体异常的比例更高。给予偶氮甲烷和DSS的转基因小鼠比给予这些药物的对照小鼠发生了更多更大的结肠息肉。芯片分析和质谱分析表明，CCAT2的表达增加了与核糖体生物发生和蛋白质合成相关的基因的表达。CCAT2 lncRNA直接与BOP1核糖体生物发生因子(BOP1)相互作用并稳定。CCAT2还增加了MYC的表达，从而激活了BOP1的表达。在结直肠癌细胞系中过表达BOP1会导致染色体错误分离，增加集落形成和侵袭力，而BOP1基因敲除则会降低存活率。BOP1通过增加调节染色体分离的极光激酶B(AURKB)的活性形式来促进CIN。与健康组织相比，在CCAT2转基因小鼠的息肉组织中，BOP1过表达。与非肿瘤组织相比，微卫星稳定型肿瘤(以CIN为特征)的CCAT2、lncRNA和BOP1mRNA或蛋白表达均增加，而在微卫星不稳定的肿瘤组织中无明显变化。CCAT2、lncRNA和BOP1基因表达水平升高与患者生存期缩短呈正相关。我们发现，CCAT2在结肠细胞中的过表达通过稳定和诱导AURKB的激活物BOP1的表达而促进CIN和癌症的发生。靶向该通路的策略可能被开发用于微卫星稳定型结直肠肿瘤的治疗。
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. 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 sulphate sodium (DSS) 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 pulldown, RNA immunoprecipitation, and SHAPE 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. 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 to organoids from control mice. The transgenic mice given azoxymethane and DSS 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 (AURKB), which regulates chromosomal segregation. BOP1 was overexpressed in polyp tissues from CCAT2 transgenic mice, compared to 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 non-tumor tissues. Increased levels of CCAT2 lncRNA and BOP1 mRNA correlated with each other and with shorter survival times of patients. We found that overexpression of CCAT2 in colon cells promotes CIN and carcinogenesis, by stabilizing and inducing expression of BOP1 an activator of AURKB. Strategies to target this pathway might be developed for treatment of patients with microsatellite stable colorectal tumors.
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影响因子：
3.9
作者：
Leroy, Bernard;Girard, Luc;Hollestelle, Antoinette;Minna, John D.;Gazdar, Adi F.;Soussi, Thierry
通讯作者：
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