标题:Characterization and functional analysis of the human microRNA let-7a2 promoter in lung cancer A549 cell lines
作者:Guan, Hengyun;Zhang, Pengju;Liu, Chang;Zhang, Ju;Huang, Zhaoqin;Chen, Weiwen;Chen, Zhaobo;Ni, Nana;Liu, Qingwei;Jiang, Anli [Aut
作者机构:[Guan, H] Institute of Biochemistry and Molecular Biology, Medical School of Shandong University, 44 Wenhua West Road, Jinan 250012, China;[ Zhang, P] 更多
通讯作者:Jiang, AL
通讯作者地址:[Jiang, AL]Shandong Univ, Sch Med, Inst Biochem & Mol Biol, 44 Wenhua W Rd, Jinan 250012, Peoples R China.
来源:Molecular biology reports
出版年:2011
卷:38
期:8
页码:5327-5334
DOI:10.1007/s11033-011-0683-8
关键词:cDNA: complementary DNA;dexamethasone: 50-02-2;microRNA;RXR;p53 protein;c-Myc;transcriptional factor;lithium chloride: 7447-41-8;Ras protein;TCF;GR;RORA protein;let-7a2 promoter: expression;upregulation;transcriptional regulation;CEBP alpha;sequence-reporter plasmid;pGL3-p7a2;MatInspector database
摘要:Recent studies have revealed that microRNAs have a strong association with cancer in humans. The miRNA let-7 is highly expressed in normal lung tissue, but frequently expressed at reduced levels in lung cancers. Let-7a2 is a member of the let-7 family. So far, little is known about the transcriptional regulation of let-7a2. Our study is focused on the characterization and functional analysis of the promoter of the human miRNA let-7a2 in A549 cell lines. Firstly, 5\' rapid amplification of cDNA ends (5\' RACE) was carried out and a 2.8 kb fragment in the upstream of let-7a2 gene was then cloned into pGL3-basic vector. Sequence analysis with the MatInspector database revealed that there were putative binding sites for some important transcriptional factors in the promoter region of let-7a2, such as p53, c-Myc, Ras, CEBP alpha, RORA, RXR, TCF, and GR. Additionally, a series of transfection and luciferase reporter assays were carried out to test let-7a2 promoter activity. RT-PCR and transfection of let-7a target sequence-reporter plasmid were performed to detect transcription levels of the let-7a2 gene in A549 cells treated with 9-cis-RA, all-trans-RA, lithium chloride or dexamethasone. Our results showed that the recombinant pGL3-p7a2 could acts as a promoter. The promoter activity of the 2.8 kb fragment could be downregulated by transfection with CEBP alpha or treatment with lithium chloride and enhanced by 9-cis-RA or all-trans-RA treatment. Furthermore, the results of RT-PCR analysis and transfection of let-7a target sequence-reporter plasmid showed that 9-cis-RA and all-trans-RA both upregulated let-7a2 expression, while lithium chloride downregulated its expression. Our results suggest that 9-cis-RA, all-trans-RA,lithium chloride and CEBP alpha might play important regulatory roles in let-7a2 gene expression in A549 cells.
收录类别:SCOPUS;SCIE
WOS核心被引频次:10
Scopus被引频次:17
资源类型:期刊论文
原文链接:https://www.scopus.com/inward/record.uri?eid=2-s2.0-84855234328&doi=10.1007%2fs11033-011-0683-8&partnerID=40&md5=b487827a227076bc5690011512580245
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