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miR-638 Suppresses Cell Proliferation in Gastric Cancer by Targeting Sp2

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Abstract

Background

MicroRNAs play important roles in the development and progression of various cancers. Recent studies have shown that miR-638 was downregulated in several tumors; however, its role in gastric cancer (GC) has not been investigated in detail.

Aims

The purpose of this study was to determine the role of miR-638 and to elucidate its regulatory mechanism in GC.

Methods

The expression levels of miR-638 and specificity protein 2 (Sp2) were detected by real-time PCR and Western blotting in GC. After pcDNA6.2-GW/EmGFP-miR-638 vector, miR-638 inhibitor and Sp2-siRNA transfection, the AGS cell proliferation was investigated by MTT assay and cell cycle, and apoptosis was detected using the Annexin V/PI. In addition, the regulation of Sp2 by miR-638 was evaluated by real-time RT-PCR, Western blot and luciferase reporter assays; cyclin D1 expression was measured by Western blotting.

Results

The expression of miR-638 is dramatically down-regulated and Sp2 expression is remarkably up-regulated in GC tissues. Luciferase assays revealed that miR-638 inhibited Sp2 expression by targeting the 3′-UTR of Sp2 mRNA. Overexpression of miR-638 and Sp2-siRNA reduced Sp2 expression at both the mRNA and protein levels in vitro, and inhibition of miR-638 increased Sp2 expression. Moreover, we found that miR-638 overexpression and Sp2-siRNA markedly suppressed cell proliferation with decreasing expression of cyclin D1 and inducing G1-phase cell-cycle arrest in vitro; inhibition of miR-638 significantly promoted cell proliferation by increasing expression of cyclin D1 and leading more cells into the S and G2/M phase.

Conclusions

Our results demonstrated that miR-638 suppressed GC cell proliferation by targeting Sp2 with influence on the expression of cyclin D1. We suggest that miR-638 might be a candidate predictor or an anticancer therapeutic target for GC patients.

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Acknowledgments

This work was supported by a grant from China Postdoctoral Science Foundation grant (No. 2013M542358) and a fund from the Fundamental Research Funds for the Central Universities, China.

Conflict of interest

None.

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Authors and Affiliations

  1. Department of Genetics and Cell Biology, Environment and Genes Related to Diseases Key Laboratory of Education Ministry, College of Medicine, Xi’an Jiaotong University, Xi’an, 710061, People’s Republic of China

    Ling Yu Zhao, Jia Han, Juan Yang, Xiao Fei Wang, Dong Dong Tong, Tu Sheng Song & Chen Huang

  2. Department of Oncology, First Affiliated Hospital of Medical College of Xi’an Jiaotong University, Xi’an, 710061, People’s Republic of China

    Yu Yao

  3. Department of Otorhinolaryngology, First Affiliated Hospital of Medical College of Xi’an Jiaotong University, Xi’an, 710061, People’s Republic of China

    Yuan Shao

Authors
  1. Ling Yu Zhao
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  2. Yu Yao
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  3. Jia Han
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  4. Juan Yang
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  5. Xiao Fei Wang
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  6. Dong Dong Tong
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  7. Tu Sheng Song
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  8. Chen Huang
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  9. Yuan Shao
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Corresponding authors

Correspondence to Chen Huang or Yuan Shao.

Additional information

Ling Yu Zhao and Yu Yao contributed equally to this work.

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Zhao, L.Y., Yao, Y., Han, J. et al. miR-638 Suppresses Cell Proliferation in Gastric Cancer by Targeting Sp2. Dig Dis Sci 59, 1743–1753 (2014). https://doi.org/10.1007/s10620-014-3087-5

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  • DOI: https://doi.org/10.1007/s10620-014-3087-5

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