Abstract

The search for anticancer MMP-9 inhibitors (MMPIs) in food products has become a major goal for research. MMPIs in soy have been related only to saponins and isoflavones, but recently, low specific protein fractions in soybeans were shown to reduce MMP-9 activity as well. The present work aimed at comparing the MMPI potential of protein fractions (P) and non-protein fractions (NP) isolated from soybean seeds, before and after soaking and cooking, mimicking dietary exposures. Reverse and substrate zymography, as well as a fluoregenic DQ gelatin assay were used to evaluate MMP-9 activities. Colon cancer cell migration and proliferation was also tested in HT29 cells. Regarding MMP-9 inhibition, proteins in soy presented IC50 values 100 times lower than non-protein extracts, and remained active after cooking, suggesting that proteins may be more effective MMP-9 inhibitors than non-protein compounds. Using the determined IC50 concentrations, NP fractions were able to induce higher inhibitions of HT29 cell migration and proliferation, but not through MMP-9 inhibition, whilst protein fractions were shown to specifically inhibit MMP-9 activity. Overall, our results show that protein fractions in soybeans might have a higher role in soy-related cancer prevention as MMPIs than previously expected. Being nontoxic and active at lower concentrations, the discovery of these heat-resistant specific MMPI proteins in soy can be of significant importance for cancer preventive diets, particularly considering the increasing use of soy proteins in food products and the controversy around isoflavones amongst consumers.

Highlights

  • Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases which are involved in the remodeling of connective tissue [1]

  • Our results show that protein fractions in soybeans might have a higher role in soy-related cancer prevention as MMP inhibitors (MMPIs) than previously expected

  • In order to use soy in MMPI-related cancer preventive diets, it becomes important to test its activities in cooked seeds

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Summary

Introduction

Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases which are involved in the remodeling of connective tissue [1]. One subgroup of MMPs called gelatinases (MMP-2 and MMP-9) has been recognized as important mediators in inflammation [2]. MMP-9 enzymes in particular are highly expressed in colorectal cancer (CRC) and pre-cancer pathologies such as ulcerative colitis and other inflammatory bowel diseases (IBDs) [8,9]. It is no surprise that in the last decade the search and development of MMP inhibitors (MMPIs), MMP-9 inhibitors, has become an important branch of research, both in academic as well as in industrial settings. Numerous MMPIs have been tested in different clinical trials [10] and were already demonstrated to be effective in reducing cancer progression/metastasis and IBD symptoms [3,8,9,11,12,13,14].

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