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/mul 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.
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 adenomapatients, and 10 healthy volunteers was detected with conventional MSP or IP-MSP. RESULTS: IP-MSP detected p16 methylation from 0.5pg/mul 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 adenomapatients and volunteers. CONCLUSIONS: IP-MSP is thus considered to be a promising procedure to detect serum methylated DNA in colorectal cancerpatients.
Authors: Maartje Massen; Kim Lommen; Kim A D Wouters; Johan Vandersmissen; Wim van Criekinge; James G Herman; Veerle Melotte; Leo J Schouten; Manon van Engeland; Kim M Smits Journal: Clin Epigenetics Date: 2022-04-27 Impact factor: 7.259
Authors: Sander Bach; Nina R Sluiter; Jamie J Beagan; Joost M Mekke; Johannes C F Ket; Nicole C T van Grieken; Renske D M Steenbergen; Bauke Ylstra; Geert Kazemier; Jurriaan B Tuynman Journal: JNCI Cancer Spectr Date: 2019-06-19