Aspirin metabolites 2,3‑DHBA and 2,5‑DHBA inhibit cancer cell growth: Implications in colorectal cancer prevention

Abstract

Although compelling evidence exists on the ability of aspirin to treat colorectal cancer (CRC), and numerous theories and targets have been proposed, a consensus has not been reached regarding its mechanism of action. In this regard, a relatively unexplored area is the role played by aspirin metabolites 2,3-dihydroxybenzoic acid (2,3-DHBA) and 2,5-dihydroxybenzoic acid (2,5-DHBA) in its chemopreventive actions. In a previous study, we demonstrated that 2,3-DHBA and 2,5-DHBA inhibited CDK1 enzyme activity in vitro. The aim of the present study was to understand the effect of these metabolites on the enzyme activity of all CDKs involved in cell cycle regulation (CDKs 1, 2, 4 and 6) as well as their effect on clonal formation in three different cancer cell lines. Additionally, in silico studies were performed to determine the potential sites of interactions of 2,3-DHBA and 2,5-DHBA with CDKs. We demonstrated that 2,3-DHBA and 2,5-DHBA inhibits CDK-1 enzyme activity beginning at 500 µM, while CDK2 and CDK4 activity was inhibited only at higher concentrations (>750 µM). 2,3-DHBA inhibited CDK6 enzyme activity from 250 µM, while 2,5-DHBA inhibited its activity >750 µM. Colony formation assays showed that 2,5-DHBA was highly effective in inhibiting clonal formation in HCT-116 and HT-29 CRC cell lines (250–500 µM), and in the MDA-MB-231 breast cancer cell line (~100 µM). In contrast 2,3-DHBA was effective only in MDA-MB-231 cells (~500 µM). Both aspirin and salicylic acid failed to inhibit all four CDKs and colony formation. Based on the present results, it is suggested that 2,3-DHBA and 2,5-DHBA may contribute to the chemopreventive properties of aspirin, possibly through the inhibition of CDKs. The present data and the proposed mechanisms should open new areas for future investigations.

Tissue-wide metabolomics reveals wide impact of gut microbiota on mice metabolite composition

“…3-(3-hydroxyphenyl)propionic acid and dihydrocaffeic acid are respectively the dehydroxylated and hydrogenated forms of caffeic acid (a common dietary component found in a variety of plant-derived food products) by gut microbiota (Bifidobacterium, Escherichia, Lactobacillus, and Clostridium) and merit attention for their antioxidant activities [103][104][105][106]. Gentisic acid, an active microbial metabolite of salicylic acid hydroxylation, is shown to inhibit colorectal cancer cell growth [52]. Some dietary phytochemicals such as ferulic/isoferulic acid and vanillic acid may be metabolized by gut microbiota.…”

Uptake Transporters of the SLC21, SLC22A, and SLC15A Families in Anticancer Therapy—Modulators of Cellular Entry or Pharmacokinetics?

“…OCTNs seem to be a factor in tumor progression; however, it remains to be investigated whether modulation of OCTN activity in cancer is a valid approach for growth repression, especially when considering their function in the organism. In this context, it seems noteworthy that there are reports linking the salicylic acid metabolites and their influence on cyclin-dependent kinases to colorectal cancer prevention by aspirin [181,182]. Moreover, a metabolomics analysis of mucosal biopsies obtained from individuals with an approximately three-year treatment with aspirin revealed an influence on carnitine shuttle metabolism [183].…”

Do Aspirin and Flavonoids Prevent Cancer through a Common Mechanism Involving Hydroxybenzoic Acids?—The Metabolite Hypothesis

“…Studies carried out in our laboratory have demonstrated that aspirin metabolites 2,3-DHBA and 2,5-DHBA are capable of inhibiting Cyclin Dependent Kinase (CDK) enzyme activity and cancer cell growth, suggesting their potential role in CRC prevention [62,63]. Our study also demonstrated that 2,5-DHBA was effective in inhibiting cell proliferation in HCT-116 and HT-29 cells [63]. It is important to note that HCT-116 cells do not express COX-2 and HT-29 cells have inactive COX-2 [64], indicating that a COX-independent mechanism is at play.…”

“…Hence, upon consumption of an 81 mg aspirin tablet, its concentration in the gut will be in the range of 0.3 mM to 1.4 mM under fed (~750 mL GI volume) and fasting (~160 mL GI volume) conditions, respectively [73]. It is important to note that not all HBAs are effective, as 4-HBA, 2,4-DHBA and 2,6-DHBA failed to inhibit cancer cell growth, suggesting that HBAs are selective in their modes of action [57,63]. It is also crucial to highlight the role of transporters in the uptake of these HBAs.…”

“…This would provide an opportunity for (i) immune surveillance, leading to the destruction of cancer cells, or (ii) DNA repair in cells containing damaged DNA, providing genetic stability, both of which are important steps in the prevention of cancer. While 2,4,6-THBA is likely to retard cell proliferation through CDK inhibition and upregulation of p21 Cip1 and p27 Kip1 , the exact mechanisms of cell growth inhibition by other HBAs is still not understood [57,63]. EGCG-Epigallocatechin gallate, C3G-Cyanidin-3-glucoside.…”