Background:
Altered expression of N-glycans such as polylactosamine is observed in colon cancer. AHL, a polylactosamine specific lectin from Adenia hondala from a medicinal plant from the Passifloraceae family, has been reported earlier.
Objective:
The aim of the present study is to study the interaction of AHL with human colon cancer epithelial HT-29 cells and colon cancer tissues.
Methods:
Cell viability was determined by MTT [3-[4, 5- dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide] assay, while cell surface binding and apoptosis by Annexin-V-PI assay. ROS production was analyzed using DCFDA [2’,7’ – dichlorofluorescindiacetate] kit method by flow cytometry. Immunohistochemistry was performed using biotinylated AHL and protein purification by affinity chromatography using asialofetuin-coupled Sepahrose -4B column.
Results:
AHL strongly binds to HT-29 cells with a Mean Fluorescence Intensity of 12.4, which could be blocked by competing for glycoprotein asialofetuin. AHL inhibits HT-29 cell growth in a dose and time-dependent manner with IC50 of 2.5µg/ml and differentially binds to human normal and cancerous tissues. AHL induces apoptosis and slight necrosis in HT-29 cells, increasing the early apoptotic population by 25.1% and 36% for 24 h and 48h, respectively, and necrotic population by 1.5% and 4.6 % at 24h and 48h, respectively, as revealed by Annexin-V-PI assay. AHL induces the release of Reactive Oxygen Species in HT-29 cells in a dose-dependent manner.
Conclusion:
To the best of knowledge, this is the first report on lectin from Adenia hondala, which is not a RIP with apoptotic and necrotic effect. These findings support the promising potential of AHL in cancer research.
Detection of Malignancy‐Associated Phosphoproteome Changes in Human Colorectal Cancer Induced by Cell Surface Binding of Growth‐Inhibitory Galectin‐4