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Molecular Medicine

, Volume 18, Issue 3, pp 389–400 | Cite as

Homeobox D10 Gene, a Candidate Tumor Suppressor, Is Downregulated through Promoter Hypermethylation and Associated with Gastric Carcinogenesis

  • Liangjing Wang
  • Shujie Chen
  • Meng Xue
  • Jing Zhong
  • Xian Wang
  • Lihong Gan
  • Emily K. Y. Lam
  • Xin Liu
  • Jianbin Zhang
  • Tianhua Zhou
  • Jun Yu
  • Hongchuan Jin
  • Jianmin Si
Research Article

Abstract

Homeobox D10 (HoxD10) gene plays a critical role in cell differentiation and morphogenesis during development. However, the function of HoxD10 in tumor progression remains largely unknown. We demonstrate that the expression of HoxD10 is commonly downregulated in gastric cancer tissues (n = 33) and cell lines (n = 8) relative to normal stomach tissues. Functionally, reexpression of HoxD10 results in significant inhibition of cell survival, induction of cell apoptosis, and impairment of cell migration and invasion. Moreover, ectopic expression of HoxD10 suppresses gastric tumor growth in a mouse xenograft model. To identify target candidates of HoxD10, we performed cDNA microarray and showed that HoxD10 regulates multiple downstream genes including IGFBP3. Reintroduction of HoxD10 transcriptionally upregulates IGFBP3, activates caspase 3 and caspase 8, and subsequently induces cell apoptosis. Methylation specific PCR revealed that HoxD10 promoter DNA was hypermethylated in gastric cancer cell lines. Additionally, 5-aza demethylation treatment could transiently reactivate the expression of HoxD10 in gastric cancer cells. HoxD10 promoter methylation frequently was detected in gastric cancer tissues obtained from endoscopic biopsies (85.7%, 24/28) and surgically resected samples (82.6%, 57/69). Intestinal metaplasia tissues showed a 60% methylation rate (18/30), but no detectable methylation in normal stomach tissues (0%, 0/10). Taken together, our results suggest that HoxD10 functions as a candidate tumor suppressor in gastric cancer, which is inactivated through promoter hypermethylation.

Notes

Acknowledgments

The project was supported by National Natural Science Foundation of China (30900676,81071961), National Basic Research Program of China (973 Program) (2012CB945004) and Science and Technology Key project of Zhejiang Province in China (2009C03012-3). The authors are grateful to Robert Weinberg of the Massachusetts Institute of Technology for kindly providing HoxD10 cDNA. We thank Lei Guo for helpful suggestions and discussion; Yan Shan, Xiaotong Hu and Fubiao Zhang for excellent technical assistance; and Manish Gala for critical review and discussion of the manuscript.

Supplementary material

10020_2012_1803389_MOESM1_ESM.pdf (887 kb)
Homeobox D10 Gene, a Candidate Tumor Suppressor, Is Downregulated through Promoter Hypermethylation and Associated with Gastric Carcinogenesis

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Authors and Affiliations

  • Liangjing Wang
    • 1
    • 2
  • Shujie Chen
    • 1
  • Meng Xue
    • 1
  • Jing Zhong
    • 2
  • Xian Wang
    • 3
  • Lihong Gan
    • 2
  • Emily K. Y. Lam
    • 4
  • Xin Liu
    • 4
  • Jianbin Zhang
    • 4
  • Tianhua Zhou
    • 5
  • Jun Yu
    • 4
  • Hongchuan Jin
    • 4
  • Jianmin Si
    • 1
  1. 1.Laboratory of Digestive DiseaseSir Run Run Shaw Clinical Medicine Institution of Zhejiang UniversityHangzhouChina
  2. 2.Department of Gastroenterology, Second Affiliated HospitalSchool of Medicine Zhejiang UniversityHangzhouChina
  3. 3.Key Laboratory of Biotherapy of Zhejiang Province, Biomedical Research Center, Sir Run Run Shaw HospitalSchool of Medicine Zhejiang UniversityHangzhouChina
  4. 4.Department of Medicine and Therapeutics, Institute of Digestive Disease, Prince of Wales HospitalThe Chinese University of Hong KongShatinHong Kong, China
  5. 5.Department of Cell Biology and Program in Molecular Cell BiologySchool of Medicine Zhejiang UniversityHangzhouChina

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