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Methylation of an intronic region regulates miR-199a in testicular tumor malignancy

Oncogene volume 30, pages 3404–3415 (2011)Cite this article

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Abstract

In the testicular cancer cell line, NT2, we previously demonstrated that differentially methylated regions were located in introns or intergenic regions, and postulated these might regulate non-coding RNAs. Three microRNAs and three small nucleolar RNAs were differentially methylated; one, miR-199a, was associated with the progression and prognosis of gastric and ovarian cancers. In this report we document, by epigenomic profiling of testicular tissue, that miR-199a is transcribed as antisense of dynamin 3 (chromosome 1q24.3), and hypermethylation of this region is correlated with miR-199a-5p/3p repression and tumor malignancy. Re-expression of miR-199a in testicular cancer cells led to suppression of cell growth, cancer migration, invasion and metastasis. The miR-199a-5p, one of two mature miRNA species derived from miR-199a, is associated with tumor malignancy. We further identified the embryonal carcinoma antigen podocalyxin-like protein 1 (PODXL), an anti-adhesive protein expressed in aggressive tumors, as a target of miR-199a-5p. We demonstrated PODXL is overexpressed in malignant testicular tumor, and cellular depletion of PODXL resulted in suppression of cancer invasion. The inverse relationship between PODXL and miR-199a-5p expression suggests PODXL is a downstream effector mediating the action of miR199a-5p. This report identifies DNA methylation, miR-199a dysregulation and PODXL as critical factors in tumor malignancy.

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Acknowledgements

We thank Dr Yun-Fai Chris Lau for providing the 833K cell line. This research was supported in part by the Intramural Research Program of the National Institutes of Health (NIH), Eunice Kennedy Shriver National Institute of Child Health and Human Development and the Chinese University of Hong Kong.

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Authors and Affiliations

  1. Section on Clinical and Developmental Genomics, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA

    H-H Cheung, A J Davis, T-L Lee, A L Pang, S Nagrani, O M Rennert & W-Y Chan

  2. School of Biomedical Sciences, Faculty of Medicine, the Chinese University of Hong Kong, Shatin, Hong Kong, SAR

    H-H Cheung & W-Y Chan

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  1. H-H Cheung
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Correspondence to W-Y Chan.

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Cheung, HH., Davis, A., Lee, TL. et al. Methylation of an intronic region regulates miR-199a in testicular tumor malignancy. Oncogene 30, 3404–3415 (2011). https://doi.org/10.1038/onc.2011.60

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