African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Full Length Research Paper

Cloning and characterization of the 5' flanking region of microRNA let-7a-1/let-7f-1 gene cluster in human lung cancer cell

Jian Zhao2#, Nana Ni1#, Chang Liu3, Pengju Zhang1, Yang Yu1, Zhaobo Chen1, Weiwen Chen1* and Anli Jiang1   #These authors contributed equally to this work.
1Institute of Biochemistry and Molecular Biology, School of Medicine, Shandong University, 44 Wenhuaxi Road, Jinan, Shandong 250012, China 2Department of Thoracic surgery, Qilu Hospital, Shandong University, Jinan, Shandong 250012, China 3R&D department, Shandong Freda Biopharm CO., LTD, Jinan, Shandong 250014, China
Email: [email protected]

  •  Accepted: 22 June 2011
  •  Published: 22 August 2011

Abstract

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.

           

Key words: let-7a-1; let-7f-1, 5’ rapid amplification of cDNA ends (5’RACE), promoter, lung cancer.

Abbreviation

ATRA, All-trans retinoic acid; 9cRA, 9-cis-retinoic acid; DEX,dexamethasone; DMSO, dimethylsulfoxide; miRNA, microRNA; RACE, rapid amplification of cDNA ends.

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This article is published under the terms of the Creative Commons Attribution License 4.0

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