PMC

Effects of BRBs on promoter methylation of SFRP2, PAX6a, p16, SFRP5, and WIF1, and on LINE-1 repetitive element in adjacent normal tissues and colorectal adenocarcinomas from 20 patients before and after 1-to-9 wks of berry treatment. *p<0.05.

Effects of BRB consumption on expression of proteins (β-catenin, E-cadherin, and c-Myc) downstream of the Wnt pathway as well as biomarkers of cell proliferation (Ki-67), apoptosis (TUNEL), and angiogenesis (CD105) in adjacent normal tissues and colorectal adenocarcinomas from 20 patients before and after treatment with BRBs for 1-to-9 wks. (A) Representative staining of β-catenin, E-cadherin, c-Myc, Ki-67, TUNEL, p16, and CD105 and (B) Dot-line plots of quantitative staining of these proteins. *p<0.05.

Effects of BRBs on promoter methylation of SFRP2, PAX6a, p16, SFRP5, and WIF1, and on DNMT1 protein expression in colorectal adenocarcinomas from 20 patients treated for an average of either 4 or 2 wks with BRBs. (A) T/H refers to adenocarcinomas taken from patients that had received an average of 85 berry doses (~4 wks). T/L refers to adenocarcinomas taken from patients that had received an average of 53 berry doses (~2 wks). Significant decreases from baseline in promoter methylation of SFRP2, PAX6a, WIF1, and all 5 genes combined are seen in T/H versus T/L groups (p<0.05). This is associated with decreased DNMT1 protein expression and longer berry treatment. (B) Heat-map of methylation changes from baseline of SFRP2, PAX6a, p16, SFRP5, and WIF1, and protein expression change of DNMT1 in colorectal adenocarcinomas.

Effects of BRBs on promoter methylation of SFRP2, PAX6a, p16, SFRP5, and WIF1, and on DNMT1 protein expression in adjacent normal tissues from 20 patients treated for an average of either 4 or 2 wks with BRBs. (A) N/H refers to adjacent normal tissue taken from patients that had received an average of 83 berry doses (~4 wks). N/L refers to adjacent normal tissue taken from patients that had received an average of 52 berry doses (~2 wks). Significant decreases from baseline in promoter methylation of SFRP2, PAX6a, and all 5 genes combined (combination of changes from SFRP2, PAX6a, p16, SFRP5, and WIF1) are seen in N/H versus N/L groups (p<0.05). This is associated with decreased DNMT1 protein expression, and longer berry treatment. (B) Heat-map of methylation changes from baseline of SFRP2, PAX6a, p16, SFRP5, and WIF1, and protein expression change of DNMT1 in adjacent normal colorectal tissues.

Effects of BRB consumption on protein expression of β-catenin, E-cadherin, and c-Myc downstream of the Wnt signaling pathway as well as biomarkers of cell proliferation (Ki-67), apoptosis (TUNEL), and angiogenesis (CD105) in (A) adjacent normal tissues and (B) adenocarcinomas from 20 colorectal cancer patients who had consumed BRBs for an average of either 4 or 2 wks. In adjacent normal tissues, nuclear β-catenin and Ki-67 staining are decreased significantly in the N/H group versus the N/L group, whereas, E-cadherin and TUNEL staining are significantly increased in N/H versus N/L. In adenocarcinomas, β-catenin, Ki-67, TUNEL, p16, and CD105 are protectively modulated to a greater extent in tumors from the T/H group than those in the T/L group.

Correlation of methylation change with DNMT1 change or total berry dose and differential responses of adenocarcinomas from the colon and rectum to BRB treatment. (A) Berry-induced methylation changes are positively correlated with changes in DNMT1 protein expression. (B) Berry-induced methylation changes are negatively correlated with total berry dose. (C) Overall, BRB treatment did not affect global methylation as measured by LINE-1 repetitive element bisulfite/pyrosequencing assay. Percent change from baseline of all 5 genes combined (SFRP2, PAX6a, p16, SFRP5, and WIF1) in adjacent normal tissues (D) and in both colon and rectal tumors (E). Colon group: combining data from descending, transverse and ascending colon. Significant differences of mean percent change in promoter methylation are observed in the following comparisons: N/H – R vs. N/L – R, T/H – R vs. T/L – R and T/L – C vs. T/L – R. C = colon, R = rectum.