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Wnt inhibitory factor 1 induces apoptosis and inhibits cervical cancer growth, invasion and angiogenesis in vivo

Oncogene volume 31, pages 2725–2737 (2012)Cite this article

Abstract

Aberrant activation of Wingless-type (Wnt)/β-catenin signaling is widespread in human cervical cancer. However, the underlying mechanisms of Wnt activation and the therapeutic potential of Wnt inhibition remain largely unknown. Here, we demonstrate that the Wnt inhibitory factor 1 (WIF1), a secreted Wnt antagonist, is downregulated in all human primary cervical tumors and cell lines analyzed. Our data reveal that WIF1 downregulation occurs due to promoter hypermethylation and is an early event in cervical oncogenesis. WIF1 re-expression upon 5-aza-2′-deoxycytidine treatment or WIF1 gene transfer induces significant apoptosis and G2/M arrest, and inhibits cervical cancer cell proliferation in vitro. Consistent with this, treatment of established mice tumor xenografts with peritumoral WIF1 gene transfer results in a significant inhibition of cancer growth and invasion. WIF1 treatment causes a significant decrease in intracellular WNT1 and TCF-4 proteins revealing novel Wnt-regulatory mechanisms. Thus, WIF1 causes a major cellular re-distribution of β-catenin and a significant inhibition of the Wnt/β-catenin pathway in tumor cells, as documented by a remarkable reversion in the expression of Wnt/β-catenin transcriptional target genes (E-cadherin, c-Myc, cyclin D1, CD44 and VEGF). Consequently, multiple critical events in tumor progression and metastasis such as cell proliferation, angiogenesis and invasion were inhibited by WIF1. In addition, WIF1 modulated the expression of specific anti-apoptotic and apoptotic proteins, thereby inducing significant apoptosis in vivo. Our findings demonstrate for the first time that WIF1 downregulation by epigenetic gene silencing is an important mechanism of Wnt activation in cervical oncogenesis. Of major clinical relevance, we show that peritumoral WIF1 gene transfer reduces not only cancer growth but also invasion in well-established tumors. Therefore, our data provide novel mechanistic insights into the role of WIF1 in cervical cancer progression, and the important preclinical validation of WIF1 as a potent drug target in cervical cancer treatment.

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Acknowledgements

We sincerely thank Dr Eric W Howard for kindly providing the pCI-blast vector. We are grateful to Dr Sripathi M Sureban and Dr Dharmalingam Subramaniam for their technical advice and graphic assistance. We also thank Mrs Geraldine A Chissoe and Mr Randal May (Advanced Immunohistochemistry and Morphology Core, OUHSC) for their technical assistance. We appreciate the helpful discussions of Dr Ileng Kumaran Ramachandran (CSHL) and Dr Sivakumar Ramadoss (UCLA). This work was supported by the Oklahoma Center for the Advancement of Science and Technology (LQ) (HR08-018). LQ holds a Presbyterian Health Foundation Endowed Chair in Otorhinolaryngology.

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Author notes

  1. S Ramalingam & S Anant

    Present address: 9Current address: Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, USA.,

  2. A Reis: Deceased: 24 December 2008.

Authors and Affiliations

  1. Department of Otorhinolaryngology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    I Ramachandran, E Thavathiru, W K Mills & L Queimado

  2. Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    S Ramalingam, G Natarajan & S Anant

  3. Department of Obstetrics and Gynecology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    D M Benbrook

  4. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    D M Benbrook

  5. Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    R Zuna & S Lightfoot

  6. Department of Dermatology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    A Reis

  7. Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    S Anant & L Queimado

  8. Department of Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    L Queimado

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  1. I Ramachandran
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Ramachandran, I., Thavathiru, E., Ramalingam, S. et al. Wnt inhibitory factor 1 induces apoptosis and inhibits cervical cancer growth, invasion and angiogenesis in vivo. Oncogene 31, 2725–2737 (2012). https://doi.org/10.1038/onc.2011.455

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