Cytoglobin, the newest member of the globin family, functions as a tumor suppressor gene

Narayan Shivapurkar, Victor Stastny, Naoki Okumura, Luc Girard, Yang Xie, Clemens Prinsen, Frederik B. Thunnissen, Ignacio I. Wistuba, Bogdan Czerniak, Eugene Frenkel, Jack A. Roth, Triantafillos Liloglou, George Xinarianos, John K. Field, John D. Minna, Adi F. Gazdar

Research output: Contribution to journalArticle

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Abstract

Cytoglobin (CYGB) is a recently discovered vertebrate globin distantly related to myoglobin with unknown function. CYGB is assigned to chromosomal region 17q25, which is frequently lost in multiple malignancies. Previous studies failed to detect evidence for mutations in the CYGB gene. Recent studies provided preliminary evidence for increased methylation of the gene in lung cancer. Our study was aimed at investigating the role of CYGB as a tumor suppressor gene. By nested methylation-specific DNA sequencing analysis of lung and breast cancer cell lines and bronchial and mammary epithelial cell lines, we identified that methylation of a 110-bp CpG-rich segment of the CYGB promoter was correlated with gene silencing. We specifically targeted this sequence and developed a quantitative methylation-specific PCR assay, suitable for high-throughput analysis. We showed that the tumor specificity of CYGB methylation in discriminating patients with and without lung cancer, using biopsies and sputum samples. We further showed the tumor specificity of this assay with multiple other epithelial and hematologic malignancies. To show tumor suppressor activity of CYGB, we performed the following: (a) RNA interference-mediated knockdown of CYGB gene on colony formation in a CYGB expression-positive lung cancer cell line, resulting in increased colony formation; (b) enforced gene expression in CYGB expression-negative lung and breast cancer cell lines, reducing colony formation; and (c) identification of potential proximate targets down-stream of the CYGB genes. Our data constitute the first direct functional evidence for CYGB, the newest member of the globin family, as a tumor suppressor gene.

Original languageEnglish (US)
Pages (from-to)7448-7456
Number of pages9
JournalCancer Research
Volume68
Issue number18
DOIs
StatePublished - Sep 15 2008

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Globins
Tumor Suppressor Genes
Methylation
Lung Neoplasms
Cell Line
cytoglobin
Neoplasms
Breast Neoplasms
Genes
Gene Knockdown Techniques
Myoglobin
Gene Silencing
Hematologic Neoplasms
RNA Interference
Sputum
DNA Sequence Analysis
Vertebrates
Breast
Epithelial Cells

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Cytoglobin, the newest member of the globin family, functions as a tumor suppressor gene. / Shivapurkar, Narayan; Stastny, Victor; Okumura, Naoki; Girard, Luc; Xie, Yang; Prinsen, Clemens; Thunnissen, Frederik B.; Wistuba, Ignacio I.; Czerniak, Bogdan; Frenkel, Eugene; Roth, Jack A.; Liloglou, Triantafillos; Xinarianos, George; Field, John K.; Minna, John D.; Gazdar, Adi F.

In: Cancer Research, Vol. 68, No. 18, 15.09.2008, p. 7448-7456.

Research output: Contribution to journalArticle

Shivapurkar, N, Stastny, V, Okumura, N, Girard, L, Xie, Y, Prinsen, C, Thunnissen, FB, Wistuba, II, Czerniak, B, Frenkel, E, Roth, JA, Liloglou, T, Xinarianos, G, Field, JK, Minna, JD & Gazdar, AF 2008, 'Cytoglobin, the newest member of the globin family, functions as a tumor suppressor gene', Cancer Research, vol. 68, no. 18, pp. 7448-7456. https://doi.org/10.1158/0008-5472.CAN-08-0565
Shivapurkar, Narayan ; Stastny, Victor ; Okumura, Naoki ; Girard, Luc ; Xie, Yang ; Prinsen, Clemens ; Thunnissen, Frederik B. ; Wistuba, Ignacio I. ; Czerniak, Bogdan ; Frenkel, Eugene ; Roth, Jack A. ; Liloglou, Triantafillos ; Xinarianos, George ; Field, John K. ; Minna, John D. ; Gazdar, Adi F. / Cytoglobin, the newest member of the globin family, functions as a tumor suppressor gene. In: Cancer Research. 2008 ; Vol. 68, No. 18. pp. 7448-7456.
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abstract = "Cytoglobin (CYGB) is a recently discovered vertebrate globin distantly related to myoglobin with unknown function. CYGB is assigned to chromosomal region 17q25, which is frequently lost in multiple malignancies. Previous studies failed to detect evidence for mutations in the CYGB gene. Recent studies provided preliminary evidence for increased methylation of the gene in lung cancer. Our study was aimed at investigating the role of CYGB as a tumor suppressor gene. By nested methylation-specific DNA sequencing analysis of lung and breast cancer cell lines and bronchial and mammary epithelial cell lines, we identified that methylation of a 110-bp CpG-rich segment of the CYGB promoter was correlated with gene silencing. We specifically targeted this sequence and developed a quantitative methylation-specific PCR assay, suitable for high-throughput analysis. We showed that the tumor specificity of CYGB methylation in discriminating patients with and without lung cancer, using biopsies and sputum samples. We further showed the tumor specificity of this assay with multiple other epithelial and hematologic malignancies. To show tumor suppressor activity of CYGB, we performed the following: (a) RNA interference-mediated knockdown of CYGB gene on colony formation in a CYGB expression-positive lung cancer cell line, resulting in increased colony formation; (b) enforced gene expression in CYGB expression-negative lung and breast cancer cell lines, reducing colony formation; and (c) identification of potential proximate targets down-stream of the CYGB genes. Our data constitute the first direct functional evidence for CYGB, the newest member of the globin family, as a tumor suppressor gene.",
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AU - Shivapurkar, Narayan

AU - Stastny, Victor

AU - Okumura, Naoki

AU - Girard, Luc

AU - Xie, Yang

AU - Prinsen, Clemens

AU - Thunnissen, Frederik B.

AU - Wistuba, Ignacio I.

AU - Czerniak, Bogdan

AU - Frenkel, Eugene

AU - Roth, Jack A.

AU - Liloglou, Triantafillos

AU - Xinarianos, George

AU - Field, John K.

AU - Minna, John D.

AU - Gazdar, Adi F.

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N2 - Cytoglobin (CYGB) is a recently discovered vertebrate globin distantly related to myoglobin with unknown function. CYGB is assigned to chromosomal region 17q25, which is frequently lost in multiple malignancies. Previous studies failed to detect evidence for mutations in the CYGB gene. Recent studies provided preliminary evidence for increased methylation of the gene in lung cancer. Our study was aimed at investigating the role of CYGB as a tumor suppressor gene. By nested methylation-specific DNA sequencing analysis of lung and breast cancer cell lines and bronchial and mammary epithelial cell lines, we identified that methylation of a 110-bp CpG-rich segment of the CYGB promoter was correlated with gene silencing. We specifically targeted this sequence and developed a quantitative methylation-specific PCR assay, suitable for high-throughput analysis. We showed that the tumor specificity of CYGB methylation in discriminating patients with and without lung cancer, using biopsies and sputum samples. We further showed the tumor specificity of this assay with multiple other epithelial and hematologic malignancies. To show tumor suppressor activity of CYGB, we performed the following: (a) RNA interference-mediated knockdown of CYGB gene on colony formation in a CYGB expression-positive lung cancer cell line, resulting in increased colony formation; (b) enforced gene expression in CYGB expression-negative lung and breast cancer cell lines, reducing colony formation; and (c) identification of potential proximate targets down-stream of the CYGB genes. Our data constitute the first direct functional evidence for CYGB, the newest member of the globin family, as a tumor suppressor gene.

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