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Connective tissue growth factor modulates oral squamous cell carcinoma invasion by activating a miR-504/FOXP1 signalling

Oncogene volume 31, pages 2401–2411 (2012)Cite this article

A Corrigendum to this article was published on 31 January 2013

Abstract

Connective tissue growth factor (CTGF) is a multi-functional secreted protein, and it has been shown either to promote or suppress tumor progression among different kinds of cancers. Here, we investigated the role of CTGF in oral squamous cell carcinoma (OSCC) invasion and metastasis. In five OSCC cell lines, endogenous CTGF negatively correlated with invasiveness. Exogenous CTGF protein or forced expression of CTGF gene in the oral cancer cell line SAS significantly decreased their invasive and migratory abilities. MicroRNA (miRNA) microarray analysis was performed in CTGF-overexpressed SAS cells (SAS/CTGF-M3) versus control cells to investigate the mechanism of CTGF-mediated inhibition of OSCC invasion. Among the miRNAs regulated by CTGF, miR-504 and miR-346 were the top two miRNAs downregulated in CTGF transfectants, and the result was confirmed by quantitative reverse transcriptase–PCR. Ectopic miR-504 increased migration and invasion in SAS/CTGF-M3, however, miR-346 did not have such impact on migration/invasion. Furthermore, we identified FOXP1, a member of forkhead transcription factors, as a target gene that takes part in the miR-504-induced cellular invasion. Knockdown of FOXP1 increased invasiveness in SAS/CTGF-M3, confirming the signal axis of CTGF/miR-504/FOXP1 in OSCC. Animal experiments showed that SAS/CTGF-M3-formed orthotopic tumors were associated with a lesser invasive phenotype than control cells. Expression of miR-504 in SAS/CTGF-M3 increased lymph node metastasis, and co-expression of FOXP1 in miR-504-transfected SAS/CTGF-M3 alleviated miR-504-induced metastasis. In OSCC samples, high CTGF was associated with a lower clinical stage and a better outcome. A reverse correlation between CTGF and miR-504, miR-504 and FOXP1, and a positive correlation between CTGF and FOXP1 were shown. Our study discovers a novel signal pathway involving the regulation of miRNA machinery by a secreted cytokine, which will be beneficial for developing therapeutic strategy against advanced OSCC.

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Acknowledgements

This work was supported by grants from National Science Council, Taiwan NSC-97-2314-B-002-031-MY2, NSC-98-2320-B-002-003 (to CCC) and NSC-99-2314-B-010-007-MY3 (to MHY).

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Authors and Affiliations

  1. Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan

    M-H Yang

  2. Division of Hematology-Oncology, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan

    M-H Yang

  3. Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan

    B-R Lin

  4. Graduate Institute of Oral Biology, School of Dentistry, National Taiwan University, Taipei, Taiwan

    C-H Chang & C-C Chang

  5. Department of Pediatrics, National Taiwan University Hospital, Taipei, Taiwan

    S-T Chen

  6. Department of Pediatrics, National Taiwan University Hospital Yun-Lin Branch, Taipei, Taiwan

    S-T Chen

  7. Graduate Institute of Clinical Dentistry, School of Dentistry, National Taiwan University, Taipei, Taiwan

    S-K Lin & M Y-P Kuo

  8. Department of Pathology, National Taiwan University Hospital, Taipei, Taiwan

    Y-M Jeng

  9. Institute of Toxicology, College of Medicine, National Taiwan University, Taipei, Taiwan

    M-L Kuo

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  1. M-H Yang
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Correspondence to C-C Chang.

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Yang, MH., Lin, BR., Chang, CH. et al. Connective tissue growth factor modulates oral squamous cell carcinoma invasion by activating a miR-504/FOXP1 signalling. Oncogene 31, 2401–2411 (2012). https://doi.org/10.1038/onc.2011.423

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