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首页> 外文期刊>African Journal of Biotechnology >Cloning and characterization of the 5 flanking region of microRNA let-7a-1/let-7f-1 gene cluster in human lung cancer cell
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Cloning and characterization of the 5 flanking region of microRNA let-7a-1/let-7f-1 gene cluster in human lung cancer cell

机译：MicroRNA Let-7a-1 / Let-7F-1基因簇中5个侧翼区域的克隆与表征人肺癌细胞

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摘要

In order to elucidate the molecular basis of microRNA?let-7a-1/let-7f-1?gene cluster, the transcription initiation site which was determined by 5’ rapid amplification of cDNA ends (5’RACE) and 2.1 kb of the 5' flanking region proximal to the?pre-let-7a-1was isolated and characterized. The promoter activity of the 2.1 kb fragment was analyzed by a firefly luciferase-encoding gene expression vector (pGL3) transiently transfected into lung cancer cell line A549. The 2.1 kb promoter of?let-7a-1/let-7f-1displayed a lower activity and was significantly enhanced by ectopic expression of c/EBPα?or p53 and treatment with?dexamethasone. Despite the induction of other let-7 family members such as?let-7a-3, let-7c?and?let-7d, all-trans retinoic acid (ATRA) and 9-cis retinoic acid (9cRA) display little enhancement effect on 2.1 kb promoter of?let-7a-1/let-7f-1, as well as 1,25-(OH)2D3.
机译：为了阐明MicroRNA的分子基础？Let-7A-1 / Let-7F-1？基因簇，通过5'CDNA末端（5'RACE）和2.1kb的快速扩增测定的转录起始位点5'侧翼区域近端θ预先置于Δ预先分离和表征。通过瞬时转染到肺癌细胞系A549中的萤火虫荧光素酶编码基因表达载体（PGL3）分析2.1kB片段的启动子活性。 2.1 kB启动子的α-7A-1 / Let-7F-1Displayed较低的活性，并且通过C /EBPα的异位表达显着增强？或p53和用α甲基塞米松治疗。尽管诱导其他让7个家族成员，如呢？Let-7A-3，Let-7C？和？Let-7D，全反式视黄酸（ATRA）和9-CIS维甲酸（9CRA）显示出几乎的增强效果在2.1 kB启动子的情况下？Let-7A-1 / Let-7F-1，以及1,25-（OH）2D3。

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