Citation
Ab Talib, N.N. and Nisha, M. and Ramasamy, Rajesh and Pang, J.C
(2024)
Preliminary studies on the effect of excretory secretory (ES) Ascaris lumbricoides antigens on colorectal cell line viability.
Tropical Biomedicine, 41 (2).
pp. 160-165.
ISSN 0127-5720; EISSN: 2521-9855
Abstract
Helminth parasites are a group of complex metazoans from various taxonomic families. Excretory
secretory (ES) by-products, secreted by living parasites from the surface, appeared to modulate the
host immunological response towards helminth infection. This study aims to investigate the effect of ES
antigen from helminth parasite on colorectal cell viability. Worm were cultured in phosphate-buffered
saline (PBS x1) at 37°C for 24 hours after being rinsed in sterile PBS. Using a mortar and pestle, the worm
was crushed vigorously using PBS. The obtained excretory secretory (ES) antigens were extracted and
filtered using a 0.22 µM filter and stored at -20°C for further assay. For LCMS, 100 µl of the extract was
analysed using Agilent ZORBAX Eclipse Plus C18 Rapid Resolution HT. The extraction of ES antigen (10
µg/ml and 20 µg/ml) was used for cell viability studies using CRC cell line HCT 116. Cell viability and
MTT assay were conducted as per the protocol mentioned in the MTT kit. The liquid chromatography
and mass spectrometry (LCMS) data indicated that the ES antigen contained metabolic compounds,
namely fatty acid, amino alcohol, indoles, sterols, glycosides, and sphingoids. For the Ascaris lumbricoides
LCMS analyses, around 405 metabolic peaks were detected. Out of which, 58 were detected via the
database were identified, while several compounds detected have anticancer properties. The MTT
assay indicated that after 24 hours and 48 hours of exposure, all treated cells showed a decrease in
cell viability compared to the control group. The preliminary studies demonstrated that the ES antigen
from Ascaris lumbricoides has some ability to decrease the cell viability of the HCT116 CRC cell line.
Further studies are needed to examine the cell cycle arrest and apoptosis effect of the ES antigen
towards the CRC cell line.
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