Microarray-based analysis: Identification of hypoxia-regulated microRNAs in retinoblastoma cells
Microarray-based analysis: Identification of hypoxia-regulated microRNAs in retinoblastoma cells
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基于微阵列的分析：鉴定视网膜母细胞瘤细胞中缺氧调节的 microRNA
DOI：
10.3892/ijo.2011.961
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发表时间：
2011-05-01
影响因子：
5.2
通讯作者：
Fan, Xianqun
中科院分区：
文献类型：
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作者：
Xu, Xiaofang;Jia, Renbing;Fan, Xianqun
低氧是视网膜母细胞瘤的一个基本特征，导致预后不良和对常规治疗的抵抗。MicroRNAs(MiRNAs)是参与细胞分化、增殖、死亡和新陈代谢等多种生物学过程的非编码小RNA。然而，视网膜母细胞瘤中缺氧与miRNAs表达的关系尚不清楚。在本研究中，我们旨在分析视网膜母细胞瘤细胞系在低氧条件下的miRNA表达模式，并确定受低氧调控的miRNAs及其可能的功能。应用基因芯片技术检测正常和低氧条件下视网膜母细胞瘤细胞(HXO-RB44)miRNA的表达谱。对差异表达的miRNAs进行生物信息学分析，以预测和分类关键的miRNAs及其靶基因。用实时荧光定量RT-PCR方法验证它们的表达。用细胞计数试剂盒检测miR-181b在RB细胞增殖中的功能意义。有46个miRNAs在低氧条件下表达变化超过2倍(34个上调，12个下调)。我们在视网膜母细胞瘤细胞中发现了包括miR-181b、miR-30c-2、miR-497和miR-491-3p在内的一组miRNAs(HRMS)，其中miR-181b是缺氧条件下最典型的差异表达miRNA。在功能上，这些HRMS参与了细胞凋亡、细胞黏附、细胞增殖和mRNA的加工，所有这些过程都与癌细胞的缺氧反应密切相关。此外，我们还发现给予miR-181b抑制剂可以抑制视网膜母细胞瘤细胞的增殖。这些发现首次证明miRNAs在视网膜母细胞瘤细胞的缺氧反应中发挥重要作用。MiR-181b，最典型的上调miRNA，可能有助于未来视网膜母细胞瘤的临床干预。
Hypoxia is an essential feature of retinoblastoma and contributes to poor prognosis and resistance to conventional therapy. MicroRNAs (miRNAs) are small non-coding RNAs involved in a wide variety of biological processes, including cell differentiation, proliferation, death and metabolism. However, the relationship between hypoxia and the expression of miRNAs in retinoblastoma is not well understood. In this study, we aimed to analyze the pattern of miRNA expression in a retinoblastoma cell line under hypoxic conditions and to identify the miRNAs regulated by hypoxia, as well as their possible functions. miRNA expression profiling in retinoblastoma cells (HXO-RB44) under normal and hypoxic conditions was assessed by microarray techniques. The differentially expressed miRNAs were subjected to bioinformatic analyses to predict and categorise the key miRNAs and their target genes. A quantitative real-time RT-PCR approach was used to validate their expression. A Cell Counting kit was used to evaluate the functional significance of miR-181b in RB cell proliferation. There were 46 miRNAs that changed expression more than 2-fold in response to hypoxia (34 up-regulated and 12 down-regulated). We identified a cluster of miRNAs that includes miR-181b, miR-30c-2, miR-497 and miR-491-3p as hypoxia-regulated miRNAs (HRMs) in retinoblastoma cells, of which miR-181b was the most typically differentially expressed miRNA under hypoxic conditions. Functionally, these HRMs are involved in apoptosis, cell adhesion, cell proliferation and mRNA processing, all processes that associate closely with the hypoxia response of cancer cells. Additionally, we found that administration of miR-181b inhibitor can suppress proliferation of retinoblastoma cells. These findings provide the first evidence that miRNAs play an important role in the hypoxia response of retinoblastoma cells. MiR-181b, the most typically up-regulated miRNA may aid in future clinical intervention of retinoblastoma.