Cytotoxic and apoptotic inducing activity of Amoora rohituka leaf extracts in human breast cancer cells

Abstract

Background: Amoora rohituka is described in Ayurveda, an Indian traditional system of medicine for management of disorders of blood, diseases of eye, helminthiasis disease, ulcer, liver disorders and splenomegaly. However, the leaves were not reported to have anticancer properties till date.

Objective: This study was carried out to evaluate the cytotoxic potential of leaf extracts of Amoora rohituka.

Materials and methods: The leaves powder was macerated in petroleum ether, ethyl acetate and methanol and evaluated their anticancer activities in vitro. The phytochemical constituents of the active (ethyl acetate) extract were screened by FTIR analysis and phytochemical screening methods.

Results: The ethyl acetate extract (RLEA) showed the presence of alkaloids, flavonoids, steroids, tannins, saponins and terpenoids. The RLEA exhibited high cytotoxic effect against human breast cancer cells, MCF-7 (IC₅₀ = 9.81 μg/mL) and induced apoptosis by altering nuclear morphology and DNA laddering. Wound healing assays explained the potency of extract to decrease the cell migration.

Conclusion: The extract of Amoora rohituka leaves exhibited anticancer activity with less toxicity and it could be used for development of alternative drugs in the treatment of human breast cancer.

Keywords: Apoptosis; Breast cancer; Cell migration; FTIR; HPTLC; Traditional medicine.

Graphical abstract

Fig. 1

Chemical Fingerprinting of A. rohituka leaf extracts in different organic solvents. Solvent System: (Toluene: Acetone: Ethyl acetate: 7.5: 2.0: 0.5); Track 1 = Petroleum Ether Extract, Track 2 = Ethyl acetate and Track 3 = Methanolic Extract; and Infrared Spectroscopy.

Fig. 2

Describing the cytotoxicity of different extracts of A. rohituka leaves with their respective IC₅₀ values (PE: Petroleum ether extract; EA: Ethyl acetate extract; M: Methanol extract).

Fig. 3

Microscopy of the cells showing alteration in their cellular and nuclear morphology in the treated group (A: treated with packlitaxel; B: treated with Amoora rohituka leave extract, RLEA; C: without any treatment, control negative) and also DNA laddering in the treated group (A: treated with packlitaxel; B: treated with RLEA; C: without any treatment, control negative) in agarose gel which supporting the induction of apoptosis by the extract.

Fig. 4

Wound healing showing inhibition of cellular migration in treated group of MCF-7 cells at the concentration of 0.78 μg/mL ethyl acetate extract of A. rohituka leaves (RLEA).