Immunoprecipitation of nucleosomal DNA is a novel procedure to improve the sensitivity of serum screening for the p16 hypermethylation associated with colon cancer

Sakamoto, Jun; Fujiya, Mikihiro; Okamoto, Kotaro; Nata, Toshie; Inaba, Yuhei; Moriichi, Kentaro; Tanabe, Hiroki; Mizukami, Yusuke; Watari, Jiro; Ashida, Toshifumi; Kohgo, Yutaka

doi:10.1016/j.canep.2010.01.004

« Previous Next »Cancer Epidemiology
Volume 34, Issue 2 , Pages 194-199, April 2010

Immunoprecipitation of nucleosomal DNA is a novel procedure to improve the sensitivity of serum screening for the p16 hypermethylation associated with colon cancer

Jun Sakamoto
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
Mikihiro Fujiya
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
- Corresponding author. Tel.: +81 166 68 2462; fax: +81 166 68 2469.
Kotaro Okamoto
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
Toshie Nata
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
Yuhei Inaba
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
Kentaro Moriichi
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
Hiroki Tanabe
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
Yusuke Mizukami
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
Jiro Watari
- Division of Upper Gastroenterology, Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Hyogo, Japan
Toshifumi Ashida
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan
Yutaka Kohgo
- Division of Gastroenterology and Hematology/Oncology, Department of Medicine, Asahikawa Medical College, 2-1-1-1, Midorigaoka-Higashi, Asahikawa, Hokkaido 078-8510, Japan

Accepted 5 January 2010. published online 20 January 2010.

Abstract

Background: We developed a novel method of methylation-specific PCR (MSP) using immunoprecipitation with anti-histone antibody (IP-MSP) to efficiently detect serum methylated DNA tightly bound to de-acetylated histones. Materials and methods: The detection limit of IP-MSP for p16 methylation was determined with a standard made by cell line (SKCO-1) lysate. p16 methylation of tumor and/or serum of 51 colorectal cancers and 10 adenoma patients, and 10 healthy volunteers was detected with conventional MSP or IP-MSP. Results: IP-MSP detected p16 methylation from 0.5pg/μl of the cell lysate. The sensitivity of IP-MSP for detecting serum p16 methylation in 27 patients with tumors characterized by p16 methylation was significantly higher than that with conventional method (81% versus 59%), particularly in Stage II patients (91% versus 45%). IP-MSP detected no p16 hypermethylation in sera of adenoma patients and volunteers. Conclusions: IP-MSP is thus considered to be a promising procedure to detect serum methylated DNA in colorectal cancer patients.