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Identification and experimental validation of G protein alpha inhibiting activity polypeptide 2 (GNAI2) as a microRNA-138 target in tongue squamous cell carcinoma

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Abstract

MicroRNA deregulation is a critical event in tumor initiation and progression. The down-regulation of microRNA-138 has been frequently observed in various cancers, including tongue squamous cell carcinoma (TSCC). Our previous studies suggest that deregulation of miR-138 is associated with the enhanced proliferation and invasion in TSCC cells. Here, we seek to identify the targets of miR-138 in TSCC, and explore their functional relevance in tumorigenesis. Our genome-wide expression profiling experiments identified a panel of 194 unique transcripts that were significantly down-regulated in TSCC cells transfected with miR-138. A comprehensive screening using six different sequence-based microRNA target prediction algorithms revealed that 51 out of these 194 down-regulated transcripts are potential direct targets for miR-138. These targets include: chloride channel, nucleotide-sensitive, 1A (CLNS1A), G protein alpha inhibiting activity polypeptide 2 (GNAI2), solute carrier family 20, member 1 (SLC20A1), eukaryotic translation initiation factor 4E binding protein 1 (EIF4EBP1), and Rho-related GTP-binding protein C (RhoC). GNAI2 is a known proto-oncogene that is involved in the initiation and progression of several different types of tumors. Direct targeting of miR-138 to two candidate binding sequences located in the 3′-untranslated region of GNAI2 mRNA was confirmed using luciferase reporter gene assays. Knockdown of miR-138 in TSCC cells enhanced the expression of GNAI2 at both mRNA and protein levels. In contrast, ectopic transfection of miR-138 reduced the expression of GNAI2, which, in consequence, led to reduced proliferation, cell cycle arrest and apoptosis. In summary, we identified a number of high-confident miR-138 target genes, including proto-oncogene GNAI2, which may play an important role in TSCC initiation and progression.

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Acknowledgments

This work was supported in part by NIH PHS grants K22DE014847, RO1CA139596, RO3CA135992, and a grant from Prevent Cancer Foundation (to X.Z.). We thank Mr. Lei Huang for his assistance on data analysis, and Ms. Katherine Long for her editorial assistance.

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Authors and Affiliations

  1. Center for Molecular Biology of Oral Diseases, College of Dentistry, University of Illinois at Chicago, 801 S. Paulina Street, Room 530C, Chicago, IL, 60612, USA

    Lu Jiang, Xiqiang Liu, Cheng Wang, Anxun Wang, Zujian Chen, Caroline E. Heidbreder & Xiaofeng Zhou

  2. Department of Bioengineering, College of Engineering, University of Illinois at Chicago, Chicago, IL, USA

    Yang Dai

  3. Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China

    Anxun Wang

  4. Department of Oral and Maxillofacial Surgery, College of Dentistry, University of Illinois at Chicago, Chicago, IL, USA

    Antonia Kolokythas

  5. Graduate College, UIC Cancer Center, University of Illinois at Chicago, Chicago, IL, USA

    Antonia Kolokythas & Xiaofeng Zhou

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  1. Lu Jiang
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  2. Yang Dai
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  3. Xiqiang Liu
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  4. Cheng Wang
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  9. Xiaofeng Zhou
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Correspondence to Xiaofeng Zhou.

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Jiang, L., Dai, Y., Liu, X. et al. Identification and experimental validation of G protein alpha inhibiting activity polypeptide 2 (GNAI2) as a microRNA-138 target in tongue squamous cell carcinoma. Hum Genet 129, 189–197 (2011). https://doi.org/10.1007/s00439-010-0915-3

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