ANTICANCER ACTIVITY OF Hordeum vulgare AND Pergularia daemia IN TRIPLE NEGATIVE BREAST CANCER CELLS
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Date
2019
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College of Veterinary and animal Science,Mannuthy
Abstract
The present study was aimed to evaluate the anticancer activity of methanol
extract of germinated seeds of Hordeum vulgare (Barley) and leaves of Pergularia
daemia (Veliparuthi) in MDA-MB-231 cell line. Qualitative phytochemical
screening showed the presence of steroids, glycosides, diterpenes, triterpenes and
saponins in H. vulgare whereas tannins and flavonoids were additionally detected
in P. daemia. The RP-HPLC analysis showed the presence of hordenine and lupeol
in H. vulgare and P. daemia respectively. In vitro antioxidant activity of the extracts
by DPPH and superoxide anion radical scavenging assays showed a significant
(p<0.05) concentration-dependent antioxidant activity for both the extracts. The
IC50 values for DPPH assay were found to be 112.75±5.74 and 100.67±7.9 μg/mL
for H. vulgare and P. daemia respectively and for superoxide radical scavenging
assay, the IC50 values were found to be 28.90±1.85 and 39.63±1.41 μg/mL for H.
vulgare and P. daemia respectively.
Cytotoxicity assay revealed that both the extracts produced potent
cytotoxicity with IC50 values of 41.28±0.30 and 35.95 35.95±3.57 μg/mL for H.
vulgare and P. daemia respectively. Morphological evaluation produced similar
cytotoxic morphological changes in cells treated with standard hordenine and
lupeol when compared with their respective extracts exhibiting morphological
alterations such as reduction in cell population, presence of apoptotic bodies, cell
shrinkage and vacuole formation. Both the extracts produced early apoptosis as
evident by acridine orange/ethidium bromide staining characterised by yellowgreen fluorescence. Hoechst staining revealed that both the extracts altered nuclear
morphology indicative of apoptosis with features such as nuclear marginalisation
and fragmentation. Mitochondrial membrane potential (MMP) assessed by JC-1
staining revealed that H. vulgare partially altered MMP whereas P. daemia
completely altered MMP. The results of the comet assay revealed that P. daemia
alone was able to cause DNA damage significantly (p<0.01). The results of western
blotting showed a significant (p<0.01) downregulation of Bcl-2 expression in P.
daemia treated cells whereas caspase-8 expression was significantly (p<0.01)
upregulated in H. vulgare treated cells. Thus, the study revealed that H. vulgare and
P. daemia produced apoptosis by extrinsic and intrinsic pathway respectively.