Cellular oxidative stress and antiproliferative effects of Cordia dichotoma (Linn.) seeds extract and their fractions on human cervix epitheloid (HeLa) and human lung (A549) carcinoma cells

Abstract

IntroductionCordia dichotoma (Linn.) is an ethnobotanically important plant species traditionally used for the treatment of several diseases in humans. The present study was aimed to explore the oxidative and anticancer properties of Cordia dichotoma seeds extract (MECD) along with its fractions against Human Cervix Epitheloid (HeLa) and human lung (A549) carcinoma cell lines. MethodsCrude extract of Cordia dichotoma seeds was prepared in methanol by continuous hot soxlation technique and the extract was concentrated to dryness. For the purpose of standardization of crude extract, HPTLC profiling of extract was done and finally, three different fractions namely chloroform, ethyl acetate, and the aqueous fraction was prepared using column chromatography. The cellular oxidative stress potential was measured in the term of intracellular reactive oxygen species (ROS) levels whereas anti-proliferative potential was evaluated by cell viability (MTT) assay, nuclear staining assays such as PI-Exclusion and Hoestch/PI double staining for the analysis of induction of apoptosis. Cellular morphology was also performed using inverted phase contrast microscope and percentage apoptosis along with apoptotic cell ratio was calculated. ResultsExposure to crude extract and their fractions in the dose ranging from 25 μg/mL to 250 μg/mL, significantly inhibited the cell viability (p < 0.001) in a dose-dependent manner. Fluorescence microscopy analysis and intracellular ROS generation exhibited significant ROS induction that might trigger the apoptotic pathways leading to the death of cancerous cells. ConclusionOur results suggest that the methanolic extract of Cordia dichotoma seeds as well as its aqueous fraction possesses maximum cytotoxicity against human cervix epitheloid (HeLa) and human lung (A549) cancer cell lines.