Long non-coding RNA HOTAIR, a driver of malignancy, predicts negative prognosis and exhibits oncogenic activity in oesophageal squamous cell carcinoma.
Long non-coding RNA HOTAIR, a driver of malignancy, predicts negative prognosis and exhibits oncogenic activity in oesophageal squamous cell carcinoma.
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长非编码 RNA HOTAIR 是恶性肿瘤的驱动因素，可预测食管鳞状细胞癌的阴性预后并表现出致癌活性
DOI：
10.1038/bjc.2013.548
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发表时间：
2013-10-15
影响因子：
8.8
通讯作者：
Han, W.
中科院分区：
文献类型：
--
作者：
Li, X.;Wu, Z.;Mei, Q.;Li, X.;Guo, M.;Fu, X.;Han, W.
HOX转录反义RNA（HOTAIR），从同源盒C基因（HOXC）基因座表达，能够重编程染色质组织和促进癌细胞转移，并可以同时结合多梳抑制复合物2，增强H3 K27三甲基化，和LSD 1-CoREST-REST复合物，这是H3 K4去甲基化的关键。在临床上，HOTAIR的过表达是多种癌症患者肿瘤进展和总生存率的有力预测因子。然而，HOTAIR和食管鳞状细胞癌（ESCC）之间的关系仍不清楚。我们研究HOTAIR在ESCC发病机制中的作用。我们使用实时定量PCR来测定ESCC细胞系和100例ESCC患者样本中HOTAIR的水平; 56例相邻非肿瘤组织用作对照。我们使用集落形成试验、锚定非依赖性生长试验、CCK-8试验、transwell迁移和侵袭试验以及膜联蛋白V结合试验来测量HOTAIR敲低和过表达在ESCC细胞系中的作用。我们分析了裸鼠ESCC异种移植瘤的生长。ESCC细胞系中基因表达和甲基化水平的变化分别使用基因表达微阵列和Infinium HumanMethylation 450 K BeadChip分析。与对照组相比，ESCC细胞系和患者样本中HOTAIR的水平增加;表达水平与疾病阶段和生存时间相关。在KYSE 510和KYSE 180 ESCC细胞系中使用小发夹RNA（shRNA）敲低HOTAIR降低了细胞在培养物中形成病灶、迁移和侵入细胞外基质的能力，改变了细胞周期进程，并增加了细胞对凋亡的敏感性。HOTAIR敲减减少了体内癌细胞转移，并且由HOTAIR沉默的ESCC细胞形成的肿瘤在大小和重量上都小于由小鼠异种移植模型中的shRNA载体对照细胞形成的肿瘤和转移。基因微阵列研究的结果表明，HOTAIR重新编程了ESCC细胞的基因表达谱，基因本体分析显示了对肿瘤发生重要的基因的富集，例如参与细胞迁移和细胞周期调控的基因。比较KYSE 180和KYSE180_HOTAIR细胞之间的基因表达谱和DNA甲基化分析显示，只有一小部分甲基化变化与基因表达变化相关。HOX转录反义RNA在ESCC细胞系和患者样品中上调，并促进小鼠ESCC细胞增殖和肿瘤转移。HOTAIR的敲低导致基因表达的显著变化，数据分析表明HOTAIR介导的基因调控在ESCC进展中具有关键作用，并且是治疗ESCC患者的新型表观遗传分子靶点。
HOX transcript antisense RNA (HOTAIR), which is expressed from the homebox C gene (HOXC) locus, is capable of reprogramming chromatin organisation and promoting cancer cell metastasis and can simultaneously bind the polycomb repressive complex 2, which enhances H3K27 trimethylation, and the LSD1-CoREST-REST complex, which is critical for H3K4 demethylation. Clinically, the overexpression of HOTAIR is a powerful predictor of the tumour progression and overall survival in patients with diverse cancers. The relationship between HOTAIR and oesophageal squamous cell carcinoma (ESCC), however, remains unclear. We investigated the role of HOTAIR in the pathogenesis of ESCC. We used quantitative real-time PCR to determine the level of HOTAIR in ESCC cell lines and 100 ESCC samples from patients; 56 adjacent non-neoplastic tissues were used as controls. We measured the effect of HOTAIR knockdown and overexpression in ESCC cell lines using colony formation assays, anchorage-independent growth assays, the CCK-8 assay, transwell migration and invasion assays, and Annexin V-binding assays. We analysed the growth of ESCC xenograft tumours in nude mice. Changes in the gene expression and methylation levels in ESCC cell lines were analysed using gene expression microarrays and the Infinium HumanMethylation450K BeadChip assay, respectively. The levels of HOTAIR were increased in ESCC cell lines and patient samples compared with the controls; the expression levels correlated with the disease stage and survival time. The knockdown of HOTAIR in the KYSE510 and KYSE180 ESCC cell lines using small hairpin RNAs (shRNAs) reduced the ability of the cells to form foci, migrate, and invade the extracellular matrix in culture, altered cell cycle progression, and increased the sensitivity of the cells to apoptosis. The HOTAIR knockdown reduced cancer cell metastasis in vivo, and the tumours formed by HOTAIR-silenced ESCC cells were smaller, both in size and weight, than the tumours and metastases formed by the shRNA vector control cells in a mouse xenograft model. The results of the gene microarray study showed that HOTAIR reprogrammed the gene expression profile of ESCC cells, and the gene ontology analysis revealed an enrichment in genes that are important for tumorigenesis, such as genes involved in cell migration and the regulation of the cell cycle. Comparing the gene expression profiles and DNA methylation analysis between the KYSE180 and KYSE180_HOTAIR cells revealed that only a small proportion of the methylation changes were correlated with gene expression changes. HOX transcript antisense RNA is upregulated in ESCC cell lines and patient samples, and promotes ESCC cell proliferation and tumour metastasis in mice. The knockdown of HOTAIR resulted in significant changes in gene expression, and data analysis suggested that HOTAIR-mediated gene regulation has a critical role in ESCC progression and is a novel epigenetic molecular target for treating ESCC patients.
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