Open AccessCytotoxic and apoptotic activity of Canavalia seed extract in HT-29 human colon carcinoma cells
Author(s) -
Geetha Suvarna,
Ashwini Prabhu,
N Katheeja Muhseena,
Bhagya B. Sharma
Publication year - 2021
Publication title -
biomedicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.106
H-Index - 9
ISSN - 0970-2067
DOI - 10.51248/.v41i1.543
Subject(s) - apoptosis , propidium iodide , cytotoxic t cell , microbiology and biotechnology , flow cytometry , cell cycle , concanavalin a , biology , cytotoxicity , mtt assay , dapi , cell growth , acridine orange , chemistry , in vitro , biochemistry , programmed cell death
and Aim: Concanavalin the lectin from Canavalia spp. share high structural similarity among the same taxon and exhibit wide range of biological functions. The antiproliferative activity of lectins ConA (C. ensiformis), ConC (C. cathartica), ConG (C. gladiata) and ConM (C. rosea) were studied in human colon carcinoma.
Materials and Methods: Human colon carcinoma HT-29 cells and human normal kidney HEK293T cells were examined. The cytotoxicity was evaluated by MTT assay. Cell apoptosis was analyzed by AO-EB (acridine orange-ethidium bromide dual staining) method. Cell cycle was investigated by flow cytometry (RNase-propidium iodide-based flow cytometry). The expression of cell signaling genes was analyzed by qRT-PCR.
Results: All concanavalins significantly inhibited the proliferation of HT-29 cells in dose-dependent manner (IC50 values were ConG-10.45, ConA-14.86, ConC-33.34 and ConM-48.98 µg/mL respectively). ConA and ConG induced apoptotic morphology in HT-29 cells without affecting HEK293T cells. Similarly, both lectins increased the sub-G0/G1 proportions in HT-29 cells dose-dependently. ConG showed down-regulation of AKT1, ERK1/2 and mutant p53 in HT-29 cells.
Conclusion: Concanavalin extract is shown to possess cytotoxic effect through inducing apoptosis, especially causing cell cycle arrest at G0/G1 phase in HT-29 cells.