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Epigenetic inactivation of T-box transcription factor 5, a novel tumor suppressor gene, is associated with colon cancer

Oncogene volume 29, pages 6464–6474 (2010)Cite this article

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Abstract

T-box transcription factor 5 (TBX5) is a member of a phylogenetically conserved family of genes involved in the regulation of developmental processes. The function of TBX5 in cancer development is largely unclear. We identified that TBX5 was preferentially methylated in cancer using methylation-sensitive arbitrarily primed PCR. We aim to clarify the epigenetic inactivation, biological function and clinical significance of TBX5 in colon cancer. Promoter methylation was evaluated by combined bisulfite restriction analysis and bisulfite genomic sequencing. Cell proliferation was examined by cell viability assay and colony formation assay, apoptosis by flow cytometry and cell migration by wound-healing assay. TBX5 target genes were identified by cDNA microarray analysis. Cox regression model and log-rank test were used to identify independent predictors of prognosis. TBX5 was silenced or downregulated in 88% (7/8) colon cancer cell lines, but was expressed in normal colon tissues. Loss of gene expression was associated with promoter methylation. The biological function of TBX5 in human colon cancer cells was examined. Re-expression of TBX5 in silenced colon cancer cell lines suppressed colony formation (P<0.001), proliferation (P<0.001), migration and induced apoptosis (P<0.01). Induction of apoptosis was mediated through cross-talk of extrinsic apoptosis pathway, apoptotic BCL2-associated X protein and Granzyme A signaling cascades. TBX5 suppressed tumor cell proliferation and metastasis through the upregulation of cyclin-dependent kinase inhibitor 2A, metastasis suppressor 1 and downregulation of synuclein gamma and metastasis-associated protein 1 family member 2. TBX5 methylation was detected in 68% (71/105) of primary colon tumors. Multivariate analysis showed that patients with TBX5 methylation had a significantly poor overall survival (P=0.0007). In conclusion, we identified a novel functional tumor suppressor gene TBX5 inactivated by promoter methylation in colon cancer. Detection of methylated TBX5 may serve as a potential biomarker for the prognosis of this malignancy.

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Abbreviations

BAX:

BCL2-associated X protein

BGS:

bisulfite genomic sequencing

CDKN2A:

cyclin-dependent kinase inhibitor 2A

CI:

confidence interval

COBRA:

combined bisulfite restriction analysis

MTA2:

metastasis-associated 1 family member 2

MTSS1:

metastasis suppressor 1

RR:

relative risks

RT–PCR:

reverse transcript–PCR

TBX5:

T-box transcription factor 5

SNCG:

synuclein gamma

TNFα:

tumor necrosis factor alpha

TNFRSF:

tumor necrosis factor receptor superfamily member

5-aza:

5-aza-2′-deoxycytidine

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Acknowledgements

This work was supported by research grant of Council Competitive Earmarked Research Grant CUHK (473008).

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Author notes

  1. X Ma: Co-first author.

Authors and Affiliations

  1. Department of Medicine and Therapeutics, Institute of Digestive Disease, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, SAR, China

    J Yu, X Ma, K F Cheung, X Li, S Wang, C W Wu, W K K Wu & J J Y Sung

  2. Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Hong Kong, SAR, China

    J Yu, X Ma, K F Cheung, X Li, S Wang, C W Wu, W K K Wu & J J Y Sung

  3. Department of Neurology, First Hospital, Hebei Medical University, Shijiazhuang, SAR, China

    X Ma & M Wang

  4. School of Public Health and Primary Care, The Chinese University of Hong Kong, Hong Kong, SAR, China

    L Tian

  5. Stanley Ho Center for Emerging Infectious Diseases, School of Public Health and Primary Care, The Chinese University of Hong Kong, Hong Kong, SAR, China

    M He

  6. Department of Surgery, The Chinese University of Hong Kong, Hong Kong, SAR, China

    S S M Ng

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Correspondence to J Yu.

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Yu, J., Ma, X., Cheung, K. et al. Epigenetic inactivation of T-box transcription factor 5, a novel tumor suppressor gene, is associated with colon cancer. Oncogene 29, 6464–6474 (2010). https://doi.org/10.1038/onc.2010.370

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