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Methylation and Gene Silencing of the Ras-Related GTPase Gene in Lung and Breast Cancers

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Abstract

Background

RRAD, a small Ras-related GTPase, is highly expressed in human skeletal muscle, lung, and heart. Although loss of expression of RRAD in breast cancer cells has been reported and it may act as an oncogene, the mechanism of silencing is unknown.

Methods

We examined (1) mRNA expression of RRAD in lung and breast cancer cell lines using RT-PCR and (2) methylation status of lung and breast cancers.

Results

Loss of RRAD expression was found in 14 of 20 (70%) NSCLC cell lines, 11 of 11 (100%) SCLC cell lines, and 8 of 10 (80%) breast cancer cell lines; expression was not affected in normal bronchial and mammary epithelial cells. Treatment of 23 expression-negative cell lines with a demethylating agent restored expression in all cases. We developed a methylation-specific assay from the analysis of bisulfite sequencing of the 5′ region of RRAD in expression-negative and positive cell lines, which resulted in good concordance between methylation and expression. Primary lung and breast cancers showed hypermethylation in 89 of 214 (42%) and 39 of 63 (62%) cases, respectively. RRAD hypermethylation correlated with smoking history and poorer prognosis in lung adenocarcinomas.

Conclusions

We conclude that epigenetic silencing of RRAD is a frequent event in lung and breast cancers, and analysis of it may provide novel opportunities for prognosis and therapy of these cancers.

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Acknowledgments

This work was supported by grants from the University of Texas Specialized Program of Research Excellence in Lung Cancer (NCI P50CA70907) and Early Detection Research Network (5U01CA8497102), the Gillson Longenbaugh Foundation, and the Emphasis Research Project by expenditure at the discretion of the president of The Chiba University in 2005.

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Authors and Affiliations

  1. Department of Thoracic Surgery, Graduate School of Medicine, Chiba University, Chiba, Japan

    Makoto Suzuki MD, Noriaki Sunaga MD, Toshihiko Iizasa MD & Takehiko Fujisawa MD

  2. Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, TX, USA

    Makoto Suzuki MD, Hisayuki Shigematsu MD, David S. Shames MD, Takao Takahashi MD, Narayan Shivapurkar PhD, John D. Minna MD & Adi F. Gazdar MD

Authors
  1. Makoto Suzuki MD
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  2. Hisayuki Shigematsu MD
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  3. David S. Shames MD
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  4. Noriaki Sunaga MD
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  5. Takao Takahashi MD
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  6. Narayan Shivapurkar PhD
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  7. Toshihiko Iizasa MD
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  8. John D. Minna MD
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  9. Takehiko Fujisawa MD
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  10. Adi F. Gazdar MD
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Corresponding author

Correspondence to Makoto Suzuki MD.

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Suzuki, M., Shigematsu, H., Shames, D.S. et al. Methylation and Gene Silencing of the Ras-Related GTPase Gene in Lung and Breast Cancers. Ann Surg Oncol 14, 1397–1404 (2007). https://doi.org/10.1245/s10434-006-9089-6

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  • DOI: https://doi.org/10.1245/s10434-006-9089-6

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