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DNA Hypermethylation of SHISA3 in Colorectal Cancer: An Independent Predictor of Poor Prognosis

  • Translational Research and Biomarkers
  • Published:
Annals of Surgical Oncology Aims and scope Submit manuscript

Abstract

Background

Shisa3 is a novel tumor suppressor identified in lung cancer. However, its antitumor activity in other human cancers and the mechanism of gene inactivation remain unknown.

Methods

SHISA3 expression was measured by reverse transcription-PCR (RT-PCR) and quantitative RT-PCR (RT-qPCR). DNA methylation was determined by bisulfite sequencing and pyrosequencing.

Results

Down-regulation of SHISA3 expression was observed in all of 11 colorectal cancer (CRC) cell lines and was further confirmed in 34 (65.4 %) of 52 colorectal carcinomas by RT-qPCR. Four of six CRC cell lines could restore SHISA3 expression after treatment with 5-aza-2′-deoxycytidine. Tumor-specific methylation of five CpG sites in the first intron of SHISA3 was identified by bisulfite sequencing, and their methylation levels were quantified in 127 pairs of primary CRC tissues by bisulfite pyrosequencing. The methylation levels of SHISA3 in tumors were noticeably higher than that in their matched normal mucosae. In addition, SHISA3 hypermethylation was significantly associated with an increased risk of disease recurrence in patients with stage II and III disease (P = 0.007) and was an independent predictor of poor overall survival [hazard ratio (HR) 2.9, 95 % confidence interval (CI) 1.5–5.8; P = 0.002] and disease-free survival (HR 4.0, 95 % CI 1.6–10.2; P = 0.003) of CRC patients.

Conclusions

SHISA3 gene is epigenetically inactivated in a substantial fraction of CRC, and its hypermethylation is of prognostic significance in predicting clinical outcome. The quantitative bisulfite pyrosequencing assay established could be a cost-effective tool for providing a potential biomarker of adverse prognosis in CRC.

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Acknowledgment

The study was supported by Grants from the National Science Council, Executive Yuan (NSC 102-2320-B-002-017), the Cardinal Tien Hospital (CTH-102-1-2A02) and the National Health Research Institutes (NHRI-EX103-10136BI), Taiwan.

Disclosure

No potential conflicts of interest exist.

Author information

Author notes

    Authors and Affiliations

    1. Department of Surgery, Cardinal Tien Hospital, New Taipei City, Taiwan

      Ming-Hong Tsai MD & Sou-Jhy Yen MD

    2. School of Medicine, Fu-Jen Catholic University, New Taipei City, Taiwan

      Ming-Hong Tsai MD

    3. Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, Taipei, Taiwan

      Wen-Chi Chen MS, Sung-Liang Yu PhD, Chun-Chieh Chen PhD, Tzu-Ming Jao PhD, Chi-Yen Huang MS, Sheng-Tai Tzeng PhD & Ya-Chien Yang PhD

    4. Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan

      Sung-Liang Yu PhD & Ya-Chien Yang PhD

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    Correspondence to Ya-Chien Yang PhD.

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    Ming-Hong Tsai and Wen-Chi Chen contributed equally to this study and share co-first authorship.

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    Tsai, MH., Chen, WC., Yu, SL. et al. DNA Hypermethylation of SHISA3 in Colorectal Cancer: An Independent Predictor of Poor Prognosis. Ann Surg Oncol 22 (Suppl 3), 1481–1489 (2015). https://doi.org/10.1245/s10434-015-4593-1

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    • DOI: https://doi.org/10.1245/s10434-015-4593-1

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