Real-Time PCR Analysis of Micro-RNA Expression in Ionizing Radiation-Treated Cells
Real-Time PCR Analysis of Micro-RNA Expression in Ionizing Radiation-Treated Cells
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DOI：
10.1089/cbr.2008.0513
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发表时间：
2009-02-01
影响因子：
3.4
通讯作者：
Chaudhry, M. Ahmad
中科院分区：
文献类型：
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作者：
Chaudhry, M. Ahmad
细胞对电离辐射的反应是非常复杂的，涉及许多途径。暴露于电离辐射后，可诱导多种生物效应，包括DNA损伤处理、信号转导、突变、基因表达改变、细胞周期阻滞、基因组不稳定、致癌诱导到细胞死亡。为了深入了解这种复杂的反应，研究人员检查了辐照细胞中基因表达的整体变化。最近的研究提供了microRNA （miRNA）与人类、细胞过程（包括癌变、细胞命运决定的时间、细胞凋亡和控制一系列事件的代谢途径）相关的证据。小的非编码miRNA正在成为控制基因表达的关键成分。由于miRNA靶向如此多的基因，我们假设它们表达的改变可能与细胞对放射治疗的总体反应有关。为了探索miRNA在细胞电离辐射应答中的作用，我们在伽玛辐射处理的Jurkat和TK6细胞中，利用茎环实时聚合酶链反应（stem-loop real-time polymerase chain reaction）监测了几种miRNA的表达水平。在电离辐射处理后，Jurkat细胞中若干功能抑制Ras癌基因mrna的let-7家族miRNA成员的表达水平上调，而在TK6细胞中则下调。与MYC易位相关的miRNA表达在两种细胞类型中均上调。还研究了miRNA在各种癌症中的调节作用。这些结果首次揭示了miRNA在辐射诱导的应激反应中的作用，并将导致对这些过程的分子细节的功能研究。
The cellular response to ionizing radiation exposure is very complex and involves many pathways. A wide variety of biologic effects are induced after exposure to ionizing radiation, ranging from DNA damage processing, signal transduction, mutations, altered gene expression, cell-cycle arrest, genomic instability, and induction of carcinogenesis to cell death. To gain insight into this complex response, global alterations in the expression of genes in irradiated cells have been examined. Recent studies have provided evidence to associate microRNA (miRNA) with man), cellular processes, including carcinogenesis, timing of cell-fate decision, apoptosis, and metabolic pathways controlling a range of events. The small noncoding miRNA are emerging as critical components in controlling the gene expression. Because miRNA target so many genes, we hypothesized that alterations in their expression may be associated with the overall response of cells to radiation treatment. To explore the role of miRNA in cellular response to ionizing radiation, we monitored the expression levels of several miRNA by employing the stem-loop real-time polymerase chain reaction in Jurkat and TK6 cells treated with gamma-radiation. The expression levels of several members of the let-7 family miRNA that functionally inhibit the mRNAs of Ras oncogenes were upregulated after ionizing radiation treatment in Jurkat cells but were down-regulated in TK6 cells. The expressions of miRNA associated with MYC translocation were upregulated in both cell types. The modulation of miRNA involved in various cancers was also examined. These results provide a first glimpse to indicate the involvement of miRNA in radiation-induced stress response and will lead to functional studies dissecting the molecular details of these processes.