Figure 1.
Comparison between levels of methylation in primary tumors vs. normal colonic mucosa.
Bisulfite pyrosequencing results for methylation of SEPT9 (A), TWIST1 (B), IGFBP3 (C), ALX4 (D), GAS7 (E) and miR137 (F) genes comparing normal mucosa from healthy controls (N–N) and primary tumors from CRC patients.
Figure 2.
Levels of methylation in normal colonic mucosa and crc tumors from all tnm stage patients.
Bisulfite pyrosequencing results of methylation of sept9 (a), twist1 (b), igfbp3 (c), alx4 (d), gas7 (e) and mir137 (f) comparing normal mucosa from healthy patients (n-n) and primary tumors from crc patients of all tnm stages i–iv. n (number of subjects); median of methylation (%).
Table 1.
Performance Characteristics Of Methylation Markers For The Identification Of Colorectal Cancer.
Figure 3.
Correlation between primary tumor methylation and age of the crc patients. linear regression analysis comparing tumor methylation of sept9 (a), twist1 (b), igfbp3 (c), alx4 (d), gas7 (e) and mir137 (f) vs age of crc patients (years).
Figure 4.
Correlation between colon mucosa methylation and age of healthy controls. linear regression analysis comparing tumor methylation of sept9 (a), twist1 (b), igfbp3 (c), alx4 (d), gas7 (e) and mir137 (f) vs age of the healthy control patients (years).
Table 2.
Molecular features associated with gene methylation in crc patients.
Figure 5.
Relationship between igfbp3 methylation and prognostic and predictive response in stage ii and iii crc patients.
(a) Disease-free survival of patients with stage ii and iii crc, according to igfbp3 methylation status (high methylation, n = 73, (30.3%) low methylation, n = 168, (69.7%). (b) Disease-free survival of patients with stage ii disease, according to igfbp3 methylation (high methylation, n = 45, (32.4%); low methylation, n = 94, (67.6%). (c) Disease-free survival of patients with stage ii and iii disease, in high igfbp3 methylation tumors based on adjuvant chemotherapy (chemotherapy, n = 20, (22.5%); no chemotherapy, n = 69, (77.5%). (d) Disease-free survival of patients with stage ii and iii disease, in low igfbp3 methylation tumors according to adjuvant chemotherapy (Chemotherapyc n = 83, (52.7%); NO Chemotherapy, n = 74, (47.2%).
Table 3.
Multivariate analysis of disease-free survival based on IGFBP3 promoter methylation in stage II and III CRC patients.
Table 4.
Multivariate analysis of disease-free survival and overall survival in stage ii and iii crc patients and igfbp3 promoter methylation status.
Figure 6.
Comparison between methylation levels within igfbp3 promoter in crc tissues.
(a) Schematic overview of the insulin-like grown factor binding protein 3 (igfbp3) gene. exons are represented by blue boxes, introns with blue lines, and the promoter region by blue dashed lines. the igfbp3 gene has one cytosine-phosphate-guanine (cpg) island at in the promoter region and exon1 underscoring the potential of modulating gene expression by means of cpg hypermethylation, and this is represented by a green box. we performed two different assays covering both cpgs islands (indicated by green arrows) using two quantitative methods: pyrosequencing and qmsp in order to correlate methylation levels of these two locations. (b) Linear regression analysis comparing igfbp3 methylation levels by pyrosequencing (% methylation) in 348 patient samples and qmsp (pmr) assays in crc tissues (n = 197).