Long Non-Coding RNA SH3PXD2A-AS1 Promotes Cell Progression Partly Through Epigenetic Silencing P57 and KLF2 in Colorectal Cancer
Long Non-Coding RNA SH3PXD2A-AS1 Promotes Cell Progression Partly Through Epigenetic Silencing P57 and KLF2 in Colorectal Cancer
复制标题
长非编码 RNA SH3PXD2A-AS1 部分通过表观遗传沉默 P57 和 KLF2 在结直肠癌中促进细胞进展
DOI：
10.1159/000489589
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Wang, Keming
中科院分区：
文献类型：
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作者：
Ma, Zhonghua;Peng, Peng;Wang, Keming
背景/目的：结直肠癌（CRC）是世界范围内最常见的恶性肿瘤之一。目前的证据已经揭示了长链非编码rna （IncRNAs）在包括CRC在内的多种癌症中的关键作用。在这项研究中，我们通过分析来自癌症基因组图谱（TCGA）和基因表达图谱（GEO）数据集的公开数据，确定了lncRNA SH3PXD2A-AS1是与CRC进展相关的新分子。方法：采用定量实时聚合酶链反应（qRT-PCR）检测SH3PXD2A-AS1在结直肠癌组织样本和结直肠癌细胞系中的表达水平。通过细胞活力检测、集落形成实验、乙基脱氧尿苷（Edu）测定和流式细胞术研究SH3PXD2A-AS1在结直肠癌增殖、细胞周期调控和细胞凋亡中的作用。Transwell法探讨SH3PXD2A-AS1对结直肠癌细胞迁移和侵袭的影响。采用裸鼠模型研究SH3PXD2A-AS1对体内肿瘤发生的影响。通过亚细胞分离、RNA免疫沉淀法（RIP）和染色质免疫沉淀法（ChIP）检测sh3pxd2a - asl介导的基因表达的分子机制。挽救试验用于确定P57和kruppel样因子2 （KLF2）是否参与sh3pxd2a - asl依赖性CRC增殖。结果：我们首先发现SH3PXD2A-AS1在结直肠癌组织和细胞系中显著上调，SH3PXD2A-AS1过表达与结直肠癌患者肿瘤大小、TNM分期、淋巴结转移相关。此外，SH3PXD2A-AS1基因敲低可抑制CRC细胞在体外的增殖、迁移和侵袭，并抑制体内的肿瘤发生。机制研究表明，SH3PXD2A-AS1可以通过与EZH2相互作用，表观遗传抑制P57和KLF2的表达。救援实验表明，sh3pxd2a - asl介导的肿瘤发生受到P57或KLF2过表达的损害。有趣的是，SH3PXD2A-AS1的表达与CRC组织样本中P57和KLF2的表达呈负相关。结论：本研究首次证实SH3PXD2A-AS1在结直肠癌中起癌基因作用，可能成为结直肠癌患者有希望的诊断或治疗靶点。
Background/Aims: Colorectal cancer (CRC) is one of the most commonly diagnosed malignancies worldwide. Current evidence has revealed the key roles of long non-coding RNAs (IncRNAs) in multiple cancers, including CRC. In this study we identified the lncRNA SH3PXD2A-AS1 as a novel molecule associated with CRC progression by analyzing the publicly available data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets. Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed to examine the expression levels of SH3PXD2A-AS1 in CRC tissue samples and CRC cell lines. Cell viability examination, colony-formation experiments, ethynyl deoxyuridine (Edu) assays and flow cytometry were performed to investigate the roles of SH3PXD2A-AS1 in CRC proliferation, cell cycle regulation, and apoptosis. Transwell assays were used to explore the effects of SH3PXD2A-AS1 on CRC cells migration and invasion. A nude mice model was used to assess the effects of SH3PXD2A-AS1 on tumorigenesis in vivo. Subcellular fractionation, RNA immunoprecipitation (RIP), and chromatin immunoprecipitation (ChIP) assays were conducted to detect the molecular mechanisms of SH3PXD2A-ASl-mediated gene expression. Rescue assays were used to determine whether P57 and Kruppel-like factor 2 (KLF2) were involved in SH3PXD2A-ASl-dependent CRC proliferation. Results: We firstly found that SH3PXD2A-AS1 was significantly upregulated in CRC tissues and cell lines, and overexpression of SH3PXD2A-AS1 was correlated with tumor size, TNM stage, and lymph node metastasis in patients with CRC. Furthermore, SH3PXD2A-AS1 knockdown inhibited CRC cells proliferation, migration and invasion in vitro, and suppressed tumorigenesis in vivo. Mechanistic studies indicated that SH3PXD2A-AS1 could epiqenetically repress P57 and KLF2 expression through interaction with EZH2. Rescue experiments suggested that SH3PXD2A-ASl-mediated oncogenesis was impaired by overexpression of P57 or KLF2. Interestingly, the expression of SH3PXD2A-AS1 was inversely correlated with the expression of P57 and KLF2 in CRC tissue samples. Conclusion: Our research presents the first evidence that SH3PXD2A-AS1 acts as an oncogene in CRC, and may be a promising diagnostic or therapeutic target in patients with CRC.