DNA hypermethyation and silencing of PITX1 correlated with advanced stage and poor postoperative prognosis of esophageal squamous cell carcinoma

Takeshi Otsubo, Kazuhiko Yamada, Teruki Hagiwara, Kenshiro Oshima, Kei Iida, Koro Nishikata, Tetsuro Toyoda, Toru Igari, Kyoko Nohara, Satoshi Yamashita, Masahira Hattori, Taeko Dohi, Yuki I. Kawamura

    Research output: Contribution to journalArticle

    6 Citations (Scopus)

    Abstract

    Esophageal squamous cell carcinoma (ESCC) is associated with the accumulation of genetic and epigenetic changes in the background mucosa. Dysregulated DNA methylation is known to lead to the inactivation of tumor suppressor genes and the activation of oncogenes. To identify the genes whose expression is perturbed by abnormal DNA methylation in ESCC, integrative transcriptomics by serial analysis of gene expression (SAGE) and methylome sequencing by methyl-DNA immunoprecipitation (MeDIP) analysis were performed. We found 159 genes with significantly decreased expression in ESCC compared to that in noncancerous esophageal mucosa. MeDIP-seq analysis identified hypermethylation in the promoter region of 56 of these genes. Using surgically resected tissues of 40 cases, we confirmed that the paired-like homeodomain 1 (PITX1) gene was hypermethylated in ESCC compared to that in normal tissues (P < 0.0001) by pyrosequencing. PITX1 overexpression in ESCC cell lines inhibited cell growth and colony formation, whereas PITX1 knockdown accelerated cell growth. A PITX1-transfected ESCC cell line, KYSE30, formed smaller tumors in nude mice than in mock-transfected cells. Hypermethylation of PITX1 was associated with tumor depth (P = 0.0011) and advanced tumor stage (P = 0.0052) and predicted poor survival in ESCC (hazard ratio, 0.1538; 95% confidence interval, 0.03159-0.7488; P = 0.0169). In this study, we found a novel tumor suppressor gene of ESCC, PITX1, which is silenced by DNA hypermethylation. Downregulation of PITX1 contributes to the growth and progression of ESCC. Hypermethylation of the PITX1 in ESCC correlated with tumor progression and advanced stage cancer, and may predict a poor prognosis.

    Original languageEnglish
    Pages (from-to)84434-84448
    Number of pages15
    JournalOncotarget
    Volume8
    Issue number48
    DOIs
    Publication statusPublished - 2017

    Fingerprint

    DNA
    DNA Methylation
    Neoplasms
    Tumor Suppressor Genes
    Immunoprecipitation
    Growth
    Esophageal Squamous Cell Carcinoma
    Genes
    Gene Expression
    Cell Line
    DNA Sequence Analysis
    Oncogenes
    Genetic Promoter Regions
    Epigenomics
    Nude Mice
    Transcriptional Activation
    Mucous Membrane
    Down-Regulation
    Confidence Intervals

    Keywords

    • Homeobox gene
    • Methylome
    • Prognosis marker
    • Transcriptome
    • Tumor suppressor gene

    ASJC Scopus subject areas

    • Oncology

    Cite this

    DNA hypermethyation and silencing of PITX1 correlated with advanced stage and poor postoperative prognosis of esophageal squamous cell carcinoma. / Otsubo, Takeshi; Yamada, Kazuhiko; Hagiwara, Teruki; Oshima, Kenshiro; Iida, Kei; Nishikata, Koro; Toyoda, Tetsuro; Igari, Toru; Nohara, Kyoko; Yamashita, Satoshi; Hattori, Masahira; Dohi, Taeko; Kawamura, Yuki I.

    In: Oncotarget, Vol. 8, No. 48, 2017, p. 84434-84448.

    Research output: Contribution to journalArticle

    Otsubo, T, Yamada, K, Hagiwara, T, Oshima, K, Iida, K, Nishikata, K, Toyoda, T, Igari, T, Nohara, K, Yamashita, S, Hattori, M, Dohi, T & Kawamura, YI 2017, 'DNA hypermethyation and silencing of PITX1 correlated with advanced stage and poor postoperative prognosis of esophageal squamous cell carcinoma' Oncotarget, vol. 8, no. 48, pp. 84434-84448. https://doi.org/10.18632/oncotarget.21375
    Otsubo, Takeshi ; Yamada, Kazuhiko ; Hagiwara, Teruki ; Oshima, Kenshiro ; Iida, Kei ; Nishikata, Koro ; Toyoda, Tetsuro ; Igari, Toru ; Nohara, Kyoko ; Yamashita, Satoshi ; Hattori, Masahira ; Dohi, Taeko ; Kawamura, Yuki I. / DNA hypermethyation and silencing of PITX1 correlated with advanced stage and poor postoperative prognosis of esophageal squamous cell carcinoma. In: Oncotarget. 2017 ; Vol. 8, No. 48. pp. 84434-84448.
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    abstract = "Esophageal squamous cell carcinoma (ESCC) is associated with the accumulation of genetic and epigenetic changes in the background mucosa. Dysregulated DNA methylation is known to lead to the inactivation of tumor suppressor genes and the activation of oncogenes. To identify the genes whose expression is perturbed by abnormal DNA methylation in ESCC, integrative transcriptomics by serial analysis of gene expression (SAGE) and methylome sequencing by methyl-DNA immunoprecipitation (MeDIP) analysis were performed. We found 159 genes with significantly decreased expression in ESCC compared to that in noncancerous esophageal mucosa. MeDIP-seq analysis identified hypermethylation in the promoter region of 56 of these genes. Using surgically resected tissues of 40 cases, we confirmed that the paired-like homeodomain 1 (PITX1) gene was hypermethylated in ESCC compared to that in normal tissues (P < 0.0001) by pyrosequencing. PITX1 overexpression in ESCC cell lines inhibited cell growth and colony formation, whereas PITX1 knockdown accelerated cell growth. A PITX1-transfected ESCC cell line, KYSE30, formed smaller tumors in nude mice than in mock-transfected cells. Hypermethylation of PITX1 was associated with tumor depth (P = 0.0011) and advanced tumor stage (P = 0.0052) and predicted poor survival in ESCC (hazard ratio, 0.1538; 95{\%} confidence interval, 0.03159-0.7488; P = 0.0169). In this study, we found a novel tumor suppressor gene of ESCC, PITX1, which is silenced by DNA hypermethylation. Downregulation of PITX1 contributes to the growth and progression of ESCC. Hypermethylation of the PITX1 in ESCC correlated with tumor progression and advanced stage cancer, and may predict a poor prognosis.",
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    AU - Oshima, Kenshiro

    AU - Iida, Kei

    AU - Nishikata, Koro

    AU - Toyoda, Tetsuro

    AU - Igari, Toru

    AU - Nohara, Kyoko

    AU - Yamashita, Satoshi

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