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Molecular cloning and preliminary analysis of the human α-methylacyl-CoA racemase promoter

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Abstract

α-Methylacyl-CoA racemase (AMACR) is an enzyme involved in β-oxidation of branched-chain fatty acids and bile acid intermediates. Recent works have revealed that AMACR is overexpressed in prostate cancer and functionally important for the growth of prostate cancer cells. Despite the recent interest in AMACR as a diagnostic marker for prostate cancer, little is known about the transcriptional regulation of AMACR in prostate cancer. To elucidate the regulation of the AMACR gene, a 2.3-kb fragment of its 5′ flanking region was cloned into pGL3-Basic, then using tansfection and Dual-luciferase reporter assay, a preliminary analysis on promoter activity and function of this 2.3-kb sequence was carried out. This 2.3-kb fragment represented promoter activity that consistent with the expression level in LNCaP and PC-3 cells respectively. Transfection experiments of 5′-deletion mutants into LNCaP cells revealed a positive-regulatory region located between nucleotides −423 and −93 that may be responsible for AMACR transactivation in LNCaP cells. Cotransfection experiments revealed that promoter activity of this 2.3-kb sequence was down-regulated by C/EBPα, p53, NF-κB p50. And data from luciferase-based reporter assays suggest that the promoter function of AMACR is independent of androgen receptor-mediated signaling.

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Acknowledgments

The authors are grateful to Dr. Young for kindly providing the pcDNA3.1-Sp1, pcDNA3.1-PTEN and pSG5-hAR, Dr. Shao for kindly providing the pcDNA3.1-PPARγ 2 and pcDNA3.1-C/EBPα, Dr. He for kindly providing the pBABEpuro-ras and pBABEhygro-myc used in this study. This work was supported by the National Natural Science Foundation supporting project (No. 30371564).

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Authors and Affiliations

  1. Institute of Biochemistry and Molecular Biology, School of Medicine, Shandong University, 44 Wenhua West Road, Jinan, 250012, People’s Republic of China

    Weiwen Chen, Weifang Wu, Chunxiao Yu, Wenwen Liu, Anli Jiang & Jianye Zhang

  2. Department of Thoracic Surgery, Qilu Hospital, Shandong University, 107 Wenhua West Road, Jinan, 250012, People’s Republic of China

    Jian Zhao

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  1. Weiwen Chen
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  2. Weifang Wu
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  3. Jian Zhao
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  4. Chunxiao Yu
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  5. Wenwen Liu
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Correspondence to Anli Jiang or Jianye Zhang.

Additional information

W. Chen and W. Wu contributed equally to the work.

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Chen, W., Wu, W., Zhao, J. et al. Molecular cloning and preliminary analysis of the human α-methylacyl-CoA racemase promoter. Mol Biol Rep 36, 423–430 (2009). https://doi.org/10.1007/s11033-007-9196-x

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  • DOI: https://doi.org/10.1007/s11033-007-9196-x

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