长链非编码RNA ATB通过负向调控miR-200a促进胶质瘤恶性进展。
Long non-coding RNA ATB promotes glioma malignancy by negatively regulating miR-200a.
作者信息
Ma Chun-Chun, Xiong Zhang, Zhu Guan-Nan, Wang Chao, Zong Gang, Wang Hong-Liang, Bian Er-Bao, Zhao Bing
机构信息
Department of Neurosurgery, The Second Affiliated Hospital of Anhui Medical University, 678 Fu Rong Road, Hefei, Anhui Province, China, 230601.
Cerebral Vascular Disease Research Center, Anhui Medical University, 678 Fu Rong Road, Hefei, Anhui Province, China, 230601.
出版信息
J Exp Clin Cancer Res. 2016 Jun 6;35(1):90. doi: 10.1186/s13046-016-0367-2.
BACKGROUND
Glioma is one of the most common and aggressive primary malignant tumor in the brain. Accumulating evidences indicated that aberrantly expressed non-coding RNAs (ncRNAs), including long non-coding RNAs (lncRNAs) and microRNAs (miRNAs), contribute to tumorigenesis. However, potential mechanisms between lncRNAs and miRNAs in glioma remain largely unknown.
METHODS
Long non-coding RNA activated by TGF-β (LncRNA-ATB) expression in glioma tissues and cells was quantified by quantitative reverse transcription-PCR. Glioma cell lines U251 and A172 were transfected with sh-ATB, miR-200a mimics, miR-200a inhibitors, after we assayed the cell phenotype and expression of the relevant molecules. Dual-luciferase reporter assay, RIP and a xenograft mouse model were used to examine the expression of sh-ATB and its target gene miR-200a.
RESULTS
ATB is abnormally up-regulated both in glioma tissues and cell lines compared with normal brain tissues, and glioma patients with high ATB expression had shorter overall survival time. Knockdown of ATB significantly inhibits glioma malignancy, including cell proliferation, colony formation, migration, invasion in vitro, and the xenograft tumor formation in vivo. In addition, ATB was confirmed to target miR-200a, and miR-200a inhibition reversed the malignant characteristics of ATB knockdown on glioma cells. In particular, ATB may act as a ceRNA, effectively becoming a sink for miR-200a, thereby modulating the derepression of TGF-β2.
CONCLUSIONS
Our findings suggest that ATB plays an oncogenic role of glioma cells by inhibiting miR-200a and facilitating TGF-β2 in glioma, thereby may represent a potential therapeutic target for the treatment of human glioma.
背景
胶质瘤是脑内最常见且侵袭性最强的原发性恶性肿瘤之一。越来越多的证据表明，异常表达的非编码RNA（ncRNAs），包括长链非编码RNA（lncRNAs）和微小RNA（miRNAs），参与肿瘤发生。然而，lncRNAs与miRNAs在胶质瘤中的潜在机制仍知之甚少。
方法
采用定量逆转录PCR法检测胶质瘤组织和细胞中转化生长因子-β激活的长链非编码RNA（LncRNA-ATB）的表达。在检测细胞表型和相关分子表达后，用sh-ATB、miR-200a模拟物、miR-200a抑制剂转染胶质瘤细胞系U251和A172。采用双荧光素酶报告基因检测、RNA免疫沉淀和异种移植小鼠模型检测sh-ATB及其靶基因miR-200a的表达。
结果
与正常脑组织相比，ATB在胶质瘤组织和细胞系中均异常上调，且ATB高表达的胶质瘤患者总生存时间较短。敲低ATB可显著抑制胶质瘤的恶性程度，包括细胞增殖、集落形成、迁移、体外侵袭以及体内异种移植瘤形成。此外，证实ATB靶向miR-200a，抑制miR-200a可逆转敲低ATB对胶质瘤细胞的恶性特征影响。特别是，ATB可能作为竞争性内源性RNA（ceRNA），有效成为miR-200a的“海绵”，从而调节转化生长因子-β2（TGF-β2）的去抑制。
结论
我们的研究结果表明，ATB通过抑制miR-200a并促进胶质瘤中TGF-β2的表达，在胶质瘤细胞中发挥致癌作用，从而可能成为治疗人类胶质瘤的潜在治疗靶点。
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