Aberrant methylation of the CDH13 (H-cadherin) promoter region in colorectal cancers and adenomas

Shinichi Toyooka, Kiyomi O. Toyooka, Kenichi Harada, Kuniharu Miyajima, Prakash Makarla, Ubaradka G. Sathyanarayana, Jing Yin, Fumiaki Sato, Narayan Shivapurkar, Stephen J. Meltzer, Adi F. Gazdar

Research output: Contribution to journalArticle

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Abstract

Expression of the cadherin family member CDH13 (H-cadherin) is reduced in several human tumors, and it has been hypothesized that this gene functions as a tumor suppressor gene. Previously, we reported that the 5′ region of CDH13 is frequently methylated in breast and lung cancers. Here we confirmed the promoter activity of 5′ region of CDH13 by luciferase assay and examined its aberrant methylation in colorectal cancers, cell lines, and adenomas. Methylation status was investigated by methylation-specific PCR (MSP) and by bisulfite DNA sequencing of cloned DNA of PCR amplicons. In cell lines, we examined the correlation between methylation status and mRNA expression by reverse transcription-PCR. Aberrant methylation of CDH13 was present in 7 of 13 (54%) cell lines, and expression was absent in 6 of 13 (46%) cell lines. CDH13 expression was present in six cell lines that showed only the unmethylated form by MSP and in one cell line that showed both the methylated and unmethylated forms. Treatment with 5-aza-2′-deoxycytidine restored CDH13 expression in methylated cell lines. In surgically resected samples, 17 of 35 (49%) cases of primary colorectal cancer, 2 of 33 (6%) cases of corresponding nonmalignant colorectal mucosa, and 8 of 19 (42%) adenomas were methylated. Sequence data after bisulfite treatment indicated that primary cancers and two cell lines with loss of expression were highly methylated compared with nonmalignant colorectal epithelial cells, especially at the attachment sites of primers for MSP, although there was heterogeneity in methylation status. Our results suggest that CDH13 expression is frequently silenced by aberrant methylation in colorectal cancers and adenomas and that methylation of CDH13 commences at an early stage of multistep colorectal tumorigenesis.

Original languageEnglish
Pages (from-to)3382-3386
Number of pages5
JournalCancer Research
Volume62
Issue number12
Publication statusPublished - Jun 15 2002
Externally publishedYes

Fingerprint

Genetic Promoter Regions
Adenoma
Methylation
Colorectal Neoplasms
Cell Line
Polymerase Chain Reaction
decitabine
H-cadherin
Cadherins
Tumor Suppressor Genes
Luciferases
DNA Sequence Analysis
Reverse Transcription
Lung Neoplasms
Neoplasms
Carcinogenesis
Mucous Membrane
Epithelial Cells
Breast Neoplasms
Messenger RNA

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Toyooka, S., Toyooka, K. O., Harada, K., Miyajima, K., Makarla, P., Sathyanarayana, U. G., ... Gazdar, A. F. (2002). Aberrant methylation of the CDH13 (H-cadherin) promoter region in colorectal cancers and adenomas. Cancer Research, 62(12), 3382-3386.

Aberrant methylation of the CDH13 (H-cadherin) promoter region in colorectal cancers and adenomas. / Toyooka, Shinichi; Toyooka, Kiyomi O.; Harada, Kenichi; Miyajima, Kuniharu; Makarla, Prakash; Sathyanarayana, Ubaradka G.; Yin, Jing; Sato, Fumiaki; Shivapurkar, Narayan; Meltzer, Stephen J.; Gazdar, Adi F.

In: Cancer Research, Vol. 62, No. 12, 15.06.2002, p. 3382-3386.

Research output: Contribution to journalArticle

Toyooka, S, Toyooka, KO, Harada, K, Miyajima, K, Makarla, P, Sathyanarayana, UG, Yin, J, Sato, F, Shivapurkar, N, Meltzer, SJ & Gazdar, AF 2002, 'Aberrant methylation of the CDH13 (H-cadherin) promoter region in colorectal cancers and adenomas', Cancer Research, vol. 62, no. 12, pp. 3382-3386.
Toyooka S, Toyooka KO, Harada K, Miyajima K, Makarla P, Sathyanarayana UG et al. Aberrant methylation of the CDH13 (H-cadherin) promoter region in colorectal cancers and adenomas. Cancer Research. 2002 Jun 15;62(12):3382-3386.
Toyooka, Shinichi ; Toyooka, Kiyomi O. ; Harada, Kenichi ; Miyajima, Kuniharu ; Makarla, Prakash ; Sathyanarayana, Ubaradka G. ; Yin, Jing ; Sato, Fumiaki ; Shivapurkar, Narayan ; Meltzer, Stephen J. ; Gazdar, Adi F. / Aberrant methylation of the CDH13 (H-cadherin) promoter region in colorectal cancers and adenomas. In: Cancer Research. 2002 ; Vol. 62, No. 12. pp. 3382-3386.
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abstract = "Expression of the cadherin family member CDH13 (H-cadherin) is reduced in several human tumors, and it has been hypothesized that this gene functions as a tumor suppressor gene. Previously, we reported that the 5′ region of CDH13 is frequently methylated in breast and lung cancers. Here we confirmed the promoter activity of 5′ region of CDH13 by luciferase assay and examined its aberrant methylation in colorectal cancers, cell lines, and adenomas. Methylation status was investigated by methylation-specific PCR (MSP) and by bisulfite DNA sequencing of cloned DNA of PCR amplicons. In cell lines, we examined the correlation between methylation status and mRNA expression by reverse transcription-PCR. Aberrant methylation of CDH13 was present in 7 of 13 (54{\%}) cell lines, and expression was absent in 6 of 13 (46{\%}) cell lines. CDH13 expression was present in six cell lines that showed only the unmethylated form by MSP and in one cell line that showed both the methylated and unmethylated forms. Treatment with 5-aza-2′-deoxycytidine restored CDH13 expression in methylated cell lines. In surgically resected samples, 17 of 35 (49{\%}) cases of primary colorectal cancer, 2 of 33 (6{\%}) cases of corresponding nonmalignant colorectal mucosa, and 8 of 19 (42{\%}) adenomas were methylated. Sequence data after bisulfite treatment indicated that primary cancers and two cell lines with loss of expression were highly methylated compared with nonmalignant colorectal epithelial cells, especially at the attachment sites of primers for MSP, although there was heterogeneity in methylation status. Our results suggest that CDH13 expression is frequently silenced by aberrant methylation in colorectal cancers and adenomas and that methylation of CDH13 commences at an early stage of multistep colorectal tumorigenesis.",
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AU - Shivapurkar, Narayan

AU - Meltzer, Stephen J.

AU - Gazdar, Adi F.

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