Background: Liver cancer is a devastating cancer with increasing incidence and mortality
rates worldwide. Plants possess numerous therapeutic properties, therefore the search for novel, naturally
occurring cytotoxic compounds is urgently needed.
Methods: The anticancer activity of plant extracts and isolated compounds from Anchusa arvensis (A.
arvensis) were studied against the cell culture of HepG-2 (human hepatocellular carcinoma cell lines)
using 3-(4,5-Dimethylthiazol-yl)-diphenyl tetrazoliumbromide (MTT) assay. Apoptosis was investigated
by performing Acridine orange –ethidium bromide staining, styox green assay and DNA interaction
study. We also used tools for computational chemistry studies of isolated compounds with the tyrosine
Results: In MTT assay, the crude extract caused a significant cytotoxic effect with IC50 of 34.14 ± 0.9
μg/ml against HepG-2 cell lines. Upon fractionation, chloroform fraction (Aa.Chm) exhibited the highest
antiproliferative activity with IC50 6.55 ± 1.2 μg/ml followed by ethyl acetate (Aa.Et) fraction (IC50,
24.59 ± 0.85 μg/ml) and n-hexane (Aa.Hex) fraction (IC50 29.53 ± 1.5μg/ml). However, the aqueous
(Aa.Aq) fraction did not show any anti-proliferative activity. Bioactivity-guided isolation led to the isolation
of two compounds which were characterized as para–methoxycatechol (1) and decane (2) through
various spectroscopic techniques. Against HepG-2 cells, compound 1 showed marked potency with IC50
6.03 ± 0.75 μg/ml followed by 2 with IC50 18.52 ± 1.9 μg/ml. DMSO was used as a negative control and
doxorubicin as a reference standard (IC50 1.3 ± 0.21 μg/ml). It was observed that compounds 1-2 caused
apoptotic cell death evaluated by Acridine orange –ethidium bromide staining, styox green assay and
DNA interaction study, therefore both compounds were tested for molecular docking studies against tyrosine
kinase to support cytotoxic activity.
Conclusion: This study revealed that the plant extracts and isolated compounds possess promising antiproliferative
activity against HepG-2 cell lines via apoptotic cell death.