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miR-21 downregulates the tumor suppressor P12CDK2AP1 and Stimulates Cell Proliferation and Invasion

Authors

  • Jun Zheng,

    1. Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi'an 710032, China
    2. Department of Biochemistry and Molecular Biology, The State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi'an 710032, China
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  • Hui Xue,

    1. Department of Orthodontics, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi'an 710032, China
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  • Tao Wang,

    1. Department of Biochemistry and Molecular Biology, The State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi'an 710032, China
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  • Yuegui Jiang,

    1. Department of Conservative Dentistry, School of Stomatology, Xi'an Jiaotong University, Xi'an 710004, China
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  • Bei Liu,

    1. Department of Biochemistry and Molecular Biology, The State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi'an 710032, China
    2. Department of Neurosurgery and Institute for Functional Brain Disorders, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China
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  • Jianhu Li,

    1. Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi'an 710032, China
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  • Yanpu Liu,

    1. Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi'an 710032, China
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  • Wei Wang,

    1. Department of Biochemistry and Molecular Biology, The State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi'an 710032, China
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  • Bin Zhang,

    Corresponding author
    1. Department of Biochemistry and Molecular Biology, The State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi'an 710032, China
    • Department of Biochemistry and Molecular Biology, The State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi'an 710032, China.
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  • Moyi Sun

    Corresponding author
    1. Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi'an 710032, China
    • Department of Oral and Maxillofacial Surgery, School of Stomatology, Fourth Military Medical University, 145 West Changle Road, Xi'an 710032, China.
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  • Jun Zheng and Hui Xue contributed equally to this work.

Abstract

The present study was undertaken to investigate the regulation of P12CDK2AP1 by miRNAs. A conserved target site for miR-21 within the CDK2AP1-3′-UTR at nt 349–370 was predicted by bioinformatics software and an inverse correlation of miR-21 and CDK2AP1 protein was observed. Highly specific amplification and quantification of miR-21 was achieved using real-time RT-PCR. Transfection of HaCaT cells with pre-miR-21 significantly suppressed a luciferase reporter including the CDK2AP1-3′-UTR, whereas transfection of Tca8113 with anti-miR-21 increased activity of this reporter. This was abolished when a construct mutated at the miR-21/nt 349–370 target site was used instead. Anti-miR-21-transfected Tca8113 cells showed an increase of CDK2AP1 protein and reduced proliferation and invasion. Resected primary tumors and tumor-free surgical margins of 18 patients with head and neck squamous cell carcinomas demonstrated an inverse correlation between miR-21 and P12CDK2AP1. This study shows that P12CDK2AP1 is downregulated by miR-21 and that miR-21 promotes proliferation and invasion in cultured cells. J. Cell. Biochem. 112: 872–880, 2011. © 2010 Wiley-Liss, Inc.

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