Differential Methylation of Dna Promoter Sequences in Peripheral Blood Mononuclear Cells As Promising Diagnostic Biomarkers for Colorectal Cancer Publisher Pubmed



M Mosallaei MEYSAM ; G Siri GOLI ; B Alani BEHRANG ; Ms Shakouri Khomartash Mehdi SHAKORI ; H Naghoosi H ; F Pourghazi FARZAD ; R Heidari REZA ; Mn Nasrollahzadeh Sabet Mehrdad NASROLLAHZADEH ; J Behroozi JAVAD
Source: Journal of Cancer Research and Therapeutics Published:2024

Abstract

Objectives: Previous reports have indicated that the methylation profile in peripheral blood mononuclear cells (PBMCs) in different genes and loci is altered in colorectal cancer (CRC). Regarding the high mortality rate and silent nature of CRC, screening and early detection can meaningfully reduce disease‑related deaths. Therefore, for the first time, we aimed to evaluate the early non‑invasive diagnosis of CRC via quantitative promoter methylation analysis of RUNX3 and RASSF1A genes in PBMCs. Materials and Methods: In the present study, we analyzed the methylation status of two important tumor suppressor genes including RUNX3 and RASSF1A in 70 CRC patients and 70 non‑malignant subjects using methylation‑quantification of endonuclease‑resistant DNA (MethyQESD), and a bisulfite conversion‑independent method. Results: RUNX3 was significantly hypermethylated in PBMCs of CRC patients compared to healthy controls (P < 0.001). By determining the efficient cutoff value, the sensitivity, and specificity of RUNX3 promoter methylation for CRC diagnosis reached 84.28% and 77.14%, respectively. The receiver operating characteristic (ROC) curve analyses demonstrated that RUNX3 promoter methylation has high accuracy (areas under the curve [AUC] = 0.840, P < 0.001) for discriminating CRC subjects from healthy individuals. Moreover, RUNX3 methylation levels in PBMCs progressively increased with the stage of the disease (P < 0.001). Although the amount of RASSF1A promoter methylation was not significantly different between CRC patients and controls as well as in different stages of the disease (P > 0.05). Conclusion: Our findings confirmed that PBMCs are reliable sources of methylation analysis for CRC screening, and RUNX3 promoter methylation can be used as a promising biomarker for early diagnosis of CRC. © 2024 Elsevier B.V., All rights reserved.