MicroRNA Expression Profiles and MiR-10a Target in Anti-benzo[a] pyrene-7, 8-diol-9, 10-epoxide-transformed Human 16HBE Cells
MicroRNA Expression Profiles and MiR-10a Target in Anti-benzo[a] pyrene-7, 8-diol-9, 10-epoxide-transformed Human 16HBE Cells
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DOI：
10.1016/s0895-3988(09)60016-7
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发表时间：
2009-02-01
影响因子：
3.5
通讯作者：
Liu, Lin-Hua
中科院分区：
文献类型：
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作者：
Shen, Yue-Lan;Jiang, Yi-Guo;Liu, Lin-Hua
目的筛选抗苯并[a]芘反式-7，8-二醇-9，10-环氧化物（anti-BPDE）诱导的人支气管上皮细胞恶性转化株16 HBE-T的miRNA谱，并分析16 HBE-T中miR-10a的靶点。方法利用一种新的微阵列平台，筛选抗BPDE转化的16 HBE-T细胞的miRNA谱。使用定量真实的时间聚合酶链反应（QRT-PCR）验证miR-10 a和miR-320的微阵列数据。通过逆转录聚合酶链反应（RT-PCR）和QRT-PCR分析miR-10a的推定靶点HOXA 1的表达。结果与对照组（16 HBE-N）相比，16 HBE-T细胞中有54个miRNAs表达差异，其中45个表达上调，9个表达下调。5种最高表达的miRNA是miR-494、miR-320、miR-498、miR-129和miR-106 a。最低表达的miRNA是miR-10a、miR-493- 5 p和miR-363*。miR-17-92簇的三个成员miR-17- 5 p、miR-20 a和miR-92在16 BHE-T中显示出与miR-21、miR-141、miR-27 a、miR-27 b、miR-16和let-7家族的miRNA相比显著更高的丰度。与16 HBE-N相比，16 HBE-T中miR-10a的推定靶HOXA 1 mRNA上调3-9倍。结论本研究提供了恶性16 HBE-T中差异表达的miRNA信息，并提示这些miRNA在抗BPDE诱导的细胞转化中可能发挥作用。HOXA 1也同样上调，表明miR-10 a与HOXA 1介导的转化过程相关。
Objective To screen miRNA profiles of malignantly transformed human bronchial epithelial cells, 16HBE-T, induced by anti-benzo[a]pyrene-trans-7,8-diol-9,10-epoxide (anti-BPDE), and to analyze putative miR-10a targets in 16HBE-T. Methods A novel microarray platform was employed to screen miRNA profiles of 16HBE-T cells transformed by anti-BPDE. Microarray data for miR-10a and miR-320 were validated using quantitative real time polymerase chain reaction (QRT-PCR). The expression of a putative target for miR-10a, HOXA1, was analyzed by reverse transcription polymerase chain reaction (RT-PCR) and QRT-PCR. Results In comparison with the vehicle-treated cells (16HBE-N), 16HBE-T exhibited differential expression of 54 miRNAs, in which, 45 were over-expressed and 9 were down-regulated. The five most highly expressed miRNAs were miR-494, miR-320, miR-498, miR-129, and miR-106a. The lowest expressed miRNAs were miR-10a, miR-493-5p, and miR-363*. Three members of miR-17-92 cluster, miR-17-5p, miR-20a, and miR-92, showed significantly higher abundance in 16BHE-T as miR-21, miR-141, miR-27a, miR-27b, miR-16 and miRNAs of the let-7 family. The putative target for miR-10a, HOXA1 mRNA was up-regulated 3-9-fold in 16HBE-T, as compared with 16HBE-N. Conclusion The findings of the study provide information on differentially expressed miRNA in malignant 16HBE-T, and also suggest a potential role of these miRNAs in cell transformation induced by anti-BPDE. HOXA1 is similarly up-regulated, suggesting that miR-10a is associated with the process of HOXA1-mediated transformation.