ABSTRACT
AIM: To assess the safety and effect of the supplementation of a patented blend of dietary phytoestrogens and insoluble fibers on estrogen receptor (ER)-ß and biological parameters in sporadic colonic adenomas. METHODS: A randomized, double-blind placebo-controlled trial was performed. Patients scheduled to undergo surveillance colonoscopy for previous sporadic colonic adenomas were identified, and 60 eligible patients were randomized to placebo or active dietary intervention (ADI) twice a day, for 60 d before surveillance colonoscopy. ADI was a mixture of 175 mg milk thistle extract, 20 mg secoisolariciresinol and 750 mg oat fiber extract. ER-ß and ER-α expression, apoptosis and proliferation (Ki-67 LI) were assessed in colon samples. RESULTS: No adverse event related to ADI was recorded. ADI administration showed a significant increases in ER-ß protein (0.822 ± 0.08 vs 0.768 ± 0.10, P = 0.04) and a general trend to an increase in ER-ß LI (39.222 ± 2.69 vs 37.708 ± 5.31, P = 0.06), ER-ß/ER-α LI ratio (6.564 ± 10.04 vs 2.437 ± 1.53, P = 0.06), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (35.592 ± 14.97 vs 31.541 ± 11.54, P = 0.07) and Ki-67 (53.923 ± 20.91 vs 44.833 ± 10.38, P = 0.07) approximating statistical significance. A significant increase of ER-ß protein (0.805 ± 0.13 vs 0.773 ± 0.13, P = 0.04), mRNA (2.278 ± 1.19 vs 1.105 ± 1.07, P < 0.02) and LI (47.533 ± 15.47 vs 34.875 ± 16.67, P < 0.05) and a decrease of ER-α protein (0.423 ± 0.06 vs 0.532 ± 0.11, P < 0.02) as well as a trend to increase of ER-ß/ER-α protein in ADI vs placebo group were observed in patients without polyps (1.734 ± 0.20 vs 1.571 ± 0.42, P = 0.07). CONCLUSION: The role of ER-ß on the control of apoptosis, and its amenability to dietary intervention, are supported in our study.
Subject(s)
Adenoma/metabolism , Colonic Neoplasms/metabolism , Colonic Polyps/metabolism , Dietary Supplements , Estrogen Receptor beta/metabolism , Phytoestrogens/chemistry , Adenoma/diagnosis , Aged , Apoptosis , Biomarkers/metabolism , Biopsy , Cell Proliferation , Colonic Neoplasms/diagnosis , Colonic Polyps/diagnosis , Colonoscopy , Diet , Double-Blind Method , Estrogen Receptor alpha/metabolism , Female , Humans , Immunohistochemistry , In Situ Nick-End Labeling , Male , Microscopy, Fluorescence , Middle AgedABSTRACT
OBJECTIVES: The APC gene mutation triggers familial adenomatous polyposis (FAP) and approximately 80% of sporadic colorectal cancers. FAP summarizes the natural history of colorectal cancer because low- and high-grade dysplastic lesions and adenocarcinoma are simultaneously present in the same patients free from individual and environmental variability factors. Estrogen receptor beta (ERß) has recently been suggested as the most likely mediator of estrogen-related anti-carcinogenic effects in Apc(Min-/+) mice and humans. In this study we assessed the ERß expression in the intestinal mucosa of FAP patients to verify its possible involvement in tumor progression in colorectal cancer. MATERIAL AND METHODS: ERß and ERα expression, cell proliferation (Ki-67) and apoptosis (TUNEL), were evaluated on archival biopsy material from six patients with FAP who underwent colectomy. RESULTS: A progressive significant decrease of ERß expression was observed in the different stages of the disease as compared to normal mucosa (p < 0.001). Interestingly, a decreased ERß expression was directly correlated with apoptosis (r = 0.76, p < 0.001), and inversely correlated with cell proliferation (r = 0.54, p < 0.05). CONCLUSIONS: ERß expression is related to the severity of the disease, supporting the role of ERß as a relevant biomarker of tumor progression and possible chemopreventive target in patients at risk of colonic neoplasia.
Subject(s)
Adenocarcinoma/genetics , Adenomatous Polyposis Coli/genetics , Colon, Descending/metabolism , Colorectal Neoplasms/genetics , DNA, Neoplasm , Estrogen Receptor beta/genetics , Gene Expression Regulation, Neoplastic , Adenocarcinoma/metabolism , Adenocarcinoma/pathology , Adenomatous Polyposis Coli/metabolism , Adenomatous Polyposis Coli/pathology , Adult , Apoptosis , Biopsy , Cell Proliferation , Colon, Descending/cytology , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/pathology , Disease Progression , Estrogen Receptor beta/biosynthesis , Female , Humans , In Situ Nick-End Labeling , Intestinal Mucosa/metabolism , Intestinal Mucosa/pathology , Male , Prognosis , Severity of Illness IndexABSTRACT
Most sporadic colorectal cancers (CRCs) develop through the adenoma-carcinoma sequence pathway and are initiated by adenomatous polyposis coli (APC) gene mutations. Estrogen receptor beta (ERbeta) is recognized to progressively reduce its expression in adenomatous and carcinomatous tissues in humans. Moreover, ERbeta deficiency enhances small intestinal tumorigenesis in rodents. In the Apc(Min/+) mouse model, we evaluated intestinal polyp development and ERbeta expression plus other biological parameters influencing tumor growth (epithelial cell proliferation, apoptosis and migration) following the addition of a combination of the ERbeta-selective agonist silymarin (SIL) and/or lignin (LIG) to a high-fat/low-fiber diet. Forty-five Apc(Min/+) mice were divided in four groups: animals fed on the tumorigenic high-fat/low-fiber diet, the tumorigenic diet supplemented with SIL (0.02%) or purified LIG (6.24%) or SIL (0.005%) + LIG (6.24%). In these animals, we assessed polyp number and volume and their degree of dysplasia together with ERbeta messenger RNA (mRNA) and protein levels and epithelial cell proliferation, migration and apoptosis. The latter group of parameters was evaluated in normal and adenomatous mucosa and the results compared with those found in wild-type (WT) mice fed on the control diet. The addition of SIL or LIG to the diet and even more the specific combination of the two significantly counteracted intestinal tumorigenesis and increased ERbeta mRNA and protein levels. Cell proliferation and apoptosis were rebalanced and cell migration accelerated, restoring values similar to those observed in WT animals. Our results further support a protective effect of ERbeta in CRC suggesting the use of the combination of SIL-LIG as a potential approach against CRC development.
Subject(s)
Adenomatous Polyposis Coli Protein/physiology , Diet , Estrogen Receptor beta/metabolism , Intestinal Neoplasms/metabolism , Intestinal Neoplasms/pathology , Adenoma/etiology , Adenoma/metabolism , Adenoma/pathology , Animals , Apoptosis , Blotting, Western , Cell Proliferation , Disease Models, Animal , Estrogen Receptor beta/genetics , Immunoenzyme Techniques , In Situ Nick-End Labeling , Intestinal Neoplasms/etiology , Intestinal Polyps/etiology , Intestinal Polyps/metabolism , Intestinal Polyps/pathology , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Up-RegulationABSTRACT
Epidemiological and experimental studies suggest a protective role of estrogens against colorectal cancer. This effect seems to be mediated by their binding to estrogen receptor beta (ER-beta), one of the two estrogen receptors with high affinity for these hormones. Very recently, the demonstration of an involvement of ER-beta in the development of adenomatous polyps of the colon has also been documented, suggesting the use of selective ER-beta agonists in primary colorectal cancer prevention. Phytoestrogens are plant-derived compounds that structurally and functionally act as estrogen-agonists in mammals. They are characterized by a higher binding affinity to ER-beta as compared to estrogen receptor alpha (ER-alpha), the other estrogen receptor subtype. These biological characteristics explain why the administration of phytoestrogens does not produce the classical side effects associated to estrogen administration (cerebro- and cardio-vascular accidents, higher incidence of endometrial and breast cancer) and makes these substances ideal candidates for the prevention of colorectal cancer.