Lathyrus sativus diamine oxidase counteracts histamine‐induced cell proliferation, migration and pro‐angiogenic mediators release in human colon adenocarcinoma cell line Caco‐2

Abstract

Because histamine is a modulator of cancer cell proliferation and invasiveness, this study aimed at investigating the effect of Lathyrus sativus–derived diamine oxidase (LSAO) and its mechanism of action on Caco‐2 cell line, considering that LSAO catalizes the oxidative deamination of histamine to the corresponding aldehyde, NH3 and H2O2. Histamine (0.01–1 μM) caused a proliferative effect on Caco‐2 cells promoting cell migration, invasion and nitric oxide and vascular endothelial growth factor release. Histamine (1 μM) stimulus also down regulated occludin expression, favouring up regulation of pro‐proliferative nuclear protein Ki67. Incubation with LSAO (0.004–0.4 μM) resulted in a significant inhibition of histamine‐induced effects. LSAO rescued occludin expression and down regulated Ki67, and it inhibited histamine‐induced increase of both MMP‐2 and 9 expression. Histamine effects were mediated by RhoA‐GTP down regulation and inversely related to phospho‐p38MAPK/p50/65 up regulation. These effects were counteracted by LSAO incubation. Histamine catabolism by LSAO accounts for a significant down regulation of proliferation and invasiveness of Caco‐2 cells. This study highlights the importance to control histamine levels in contrasting pro‐angiogenic and metastatization capability of colon cancer cells and expands the knowledge about the diamine oxidase from L. sativus seeding as a phytotherapeutic approach for colon cancer.

Activation of P2×7 Receptor Promotes the Invasion and Migration of Colon Cancer Cells via the STAT3 Signaling

“…Moreover, MMP-2 plays an role in the migration and invasion of tumor cells (Ni et al, 2019). Different studies have shown that MMP-2 contributes to the migration and metastasis of colon cancer cells (Ko et al, 2019;Pietrangeli et al, 2019). Studies have shown that increased expression of P2 × 7 receptor can increase the expression of MMP-2 and MMP-9 through NF-kB, ERK1/2 and Akt signals, and promote the migration of prostate cancer and breast cancer cells (Tafani et al, 2011).…”

Lathyrus sativus diamine oxidase reduces Clostridium difficile toxin A‐induced toxicity in Caco‐2 cells by rescuing RhoA‐GTPase and inhibiting pp38‐MAPK/NF‐κB/HIF‐1α activation

“…LSAO was purified from L. sativus seedlings as previously described (Pietrangeli et al, 2019). The enzyme was characterized for protein content and enzymatic activity as reported (Pietrangeli, Federico, Mondovì, & Morpurgo, 2007), then aliquoted and stored at −20°C.…”

“…Control group cells were treated with pyrogen‐free sterile vehicle. TcdA and LSAO concentrations were selected according to our previous research studies (Esposito et al, 2016; Pietrangeli et al, 2019). For each experiment, LSAO treatment for 24 hr preceded TcdA challenge.…”

“…Absorbance was measured on a microtiter plate reader. TNF‐α, IL‐6, and VEGF levels were thus determined using the standard curves method as described elsewhere (Han, Song, & Zhang, 2016; Pietrangeli et al, 2019).…”

“…Animal and vegetal DAO food supplements resulted effective in the treatment of various gastrointestinal histaminosis and intestine inflammatory conditions like Crohn's disease and ulcerative colitis, respectively (Calinescu et al, 2012). Beyond its effect on histamine metabolism, we have recently demonstrated that LSAO markedly inhibits the release of pro‐inflammatory and angiogenic factors, such as vascular endothelial growth factor (VEGF) and nitric oxide (NO), and prevents histamine‐induced damage in human colon epithelial cell line (Caco‐2) through the rescue of RhoA‐GTPase expression and phospho‐p38MAPK/p50/65 up‐regulation (Pietrangeli et al, 2019). Here, we tested the hypothesis that LSAO also exerts a protective role in TcdA‐induced epithelial damage using the Caco‐2 cell line as a model for the human intestinal barrier, and we explored its possible mechanism of action.…”