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A Polylactosamine Specific Lectin from Adenia hondala Induces Apoptosis and Necrosis in Human Epithelial Colon Cancer HT-29 Cells

Author(s):

Shashikala R. Inamdar*, Narasimhappagari Jagadeesh, Kavita Y. Hiremath, Shivakumar Belur and Mamta Sharma   Pages 1 - 7 ( 7 )

Abstract:


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, apoptosis by Annexin-V-PI assay and ROS production using DCFDA [2’,7’ - dichlorofluorescindiacetate] kit method were analysed by flowcytometry, immunohistochemistry was performed using biotinylated AHL, 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 glycoprotein asialofetuin. AHL inhibits HT-29 cell growth in a dose and time-dependent manner with IC50 of 2.5 μg/mL and differentially binds to human normal and cancerous tissues. AHL induces apoptosis and slight necrosis in HT-29 cells with an increase in the early apoptotic population of 25.1 and 36% for 24 h and 48 h respectively and necrotic population of 1.5 and 4.6% at 24 h and 48 h 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 effects. These findings support the promising potential of AHL in cancer research.

Keywords:

AHL, apoptosis, necrosis, HT-29 cells, histochemistry, ROS.

Affiliation:

Department of Studies in Biochemistry, Karnatak University, Dharwad-580003, Department of Studies in Biochemistry, Karnatak University, Dharwad-580003, Department of Studies in Biochemistry, Karnatak University, Dharwad-580003, Department of Studies in Biochemistry, Karnatak University, Dharwad-580003, Department of Studies in Biochemistry, Karnatak University, Dharwad-580003



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