Effects of calcium and vitamin D3 on transforming growth factors in rectal mucosa of sporadic colorectal adenoma patients: a randomized controlled trial

Abstract

Transforming growth factor alpha (TGFα) and TGFβ1 are growth-promoting and -inhibiting autocrine/paracrine growth factors, respectively, that may (1) affect risk for colorectal cancer and (2) be modifiable by anti-proliferative exposures. The effects of supplemental calcium and vitamin D3 on these two markers in the normal-appearing colorectal mucosa in humans are unknown. We conducted a pilot, randomized, double-blind, placebo-controlled, 2 × 2 factorial clinical trial (n = 92; 23/treatment group) of calcium 2 g and/or vitamin D3 800 IU/d versus placebo over 6 mo. TGFα and TGFβ1 expression was measured in biopsies of normal-appearing rectal mucosa using automated immunohistochemistry and quantitative image analysis at baseline and 6-mo follow-up. In the calcium, vitamin D3 , and calcium plus vitamin D3 groups relative to the placebo group (1) the mean overall expression of TGFβ1 increased by 14% (P= 0.25), 19% (P = 0.17), and 22% (P = 0.09); (2) the ratio of TGFα expression in the upper 40% (differentiation zone) to that in the lower 60 (proliferation zone) of the crypts decreased by 34% (P = 0.11), 31% (P = 0.22), and 26% (P = 0.33); and (3) the TGFα/TGFβ1 ratio in the upper 40% of the crypts decreased by 28% (P = 0.09), 14% (P = 0.41), and 22% (P = 0.24), respectively. These preliminary results, although not statistically significant, suggest that supplemental calcium and vitamin D3 may increase TGFβ1 expression and shift TGFα expression downward from the differentiation to the proliferation zone in the crypts in the normal-appearing colorectal mucosa of sporadic colorectal adenoma patients, and support further investigation in a larger clinical trial.

Trial registration: ClinicalTrials.gov NCT00208793.

Keywords: calcium; colorectal neoplasms; transforming growth factor alpha (TGFα); transforming growth factor beta 1 (TGFβ1); vitamin D3.

© 2013 Wiley Periodicals, Inc.

Figures

Figure 1

Process of quantitative image analysis. (A) Identifying scorable crypts; (B) tracing the hemicrypt; (C) automated sectioning of the trace; and (D) automated quantification of TGFα labeling optical density in the whole hemicrypt and each section.

Figure 2

Standardized transforming growth factor alpha (TGFα) expression (labeling optical density) along normal colorectal crypts by treatment group (A - Placebo group, B - Calcium group, C - Vitamin D3 group, D -Calcium plus Vitamin D3 group) at baseline and 6-mo follow-up. The distributions were modeled and graphically plotted using the LOESS procedure.

Figure 3

Standardized transforming growth factor beta 1 (TGFβ1) expression (labeling optical density) along normal colorectal crypts by treatment group (A - Placebo group, B - Calcium group, C - Vitamin D3 group, D -Calcium plus Vitamin D3 group) at baseline and 6-mo follow-up. The distributions were modeled and graphically plotted using the LOESS procedure.