Acovenoside A Induces Mitotic Catastrophe Followed by Apoptosis in Non-Small-Cell Lung Cancer Cells

Abstract

We investigated the cytotoxic potential of the cardenolide glycoside acovenoside A against non-small-cell lung cancer cells. Lung cancer is the leading cause of cancer-related mortality and the second most common cancer diagnosed. Epidemiological studies revealed a direct correlation between the regular administration of cardiac glycosides and a lower incidence of various cancers. Acovenoside A, isolated from the pericarps of Acokanthera oppositifolia, potently inhibited proliferation and induced cytotoxicity in A549 non-small-cell lung cancer cells with an IC of 68 ± 3 nM after 48 h of exposure. Compared to the antineoplastic agent doxorubicin, acovenoside A was more potent in inhibiting the viability of A549 cancer cells. Moreover, acovenoside A exhibited selectivity against cancer cells, being significantly less toxic to lung fibroblasts and nontoxic for peripheral blood mononuclear cells. Analysis of the cell cycle profile in acovenoside A-treated A549 cells revealed mitotic arrest, due to accumulation of the G/M regulators cyclin B and CDK1, and cytokinesis failure. Furthermore, acovenoside A affected the mitochondrial membrane integrity and induced production of radical oxygen species, which resulted in induction of canonical apoptosis, manifested by caspase 3 activation and DNA fragmentation. Based on our results, acovenoside A warrants further exploration as a potential anticancer lead.

Chrysosplenol d, a Flavonol from Artemisia annua, Induces ERK1/2-Mediated Apoptosis in Triple Negative Human Breast Cancer Cells

“…Cells were cultured according to the supplier s recommendations. PBMC were isolated from whole blood of healthy donors by density gradient centrifugation using Biocoll separating solution (Biochrom GmbH, Berlin, Germany) as previously described [37,38]. The collection and analysis of peripheral blood mononuclear cells used in this study was approved by the institutional Ethics Committee (# 177/18).…”

A Novel Polyhalogenated Monoterpene Induces Cell Cycle Arrest and Apoptosis in Breast Cancer Cells

“…The cell viability assay is based on the measurement of the optical density of the orange colored water-soluble formazan salt produced by viable cells with active mitochondrial dehydrogenase (mitochondrial reduction of tetrazolium salt). Cells were plated in 96-well microtiter plates and treated with the respective compounds for the specified times before incubating them with XTT labeling mixture at 37 °C [36,37,38]. The spectrophotometric absorbance was measured using a TECAN Infinite ® 200 PRO microplate reader (Männedorf, Switzerland) at 450 nm with a 630 nm-reference filter.…”

“…Caspase 3 activity was analyzed using fluorometric detection of the cleaved tetrapeptide caspase 3/7 substrate Z-DEVD-R110. Briefly, the collected MDA-MB-231 cells were incubated with caspase 3/7 substrate (Z-DEVD-R110, 100 µM) in PBS for 1 h at 37 °C [36,37] and analyzed by flow cytometry (FACS Verse). To validate the contribution of caspase-dependent apoptosis in PPM1-induced cytotoxicity, MDA-MB-231 cells were pretreated with pancaspase inhibitor (z-VAD-fmk, 50 µM) (Calbiochem, San Diego, CA, USA) for 2 h before treatment with PPM1 (10 µM) for further 48 h and cell viability was analyzed using the XTT cell viability assay.…”

“…By contrast, the monomeric form of the dye found in the cytosol of the cells upon leakage from damaged mitochondria emits green fluorescence at 527 nm, when excited at 490 nm. Treated cells were loaded with 10 µg/mL JC-1 dye in serum-free medium for 20 min at 37 °C [36,37] and analyzed by flow cytometry (FACS Verse, BD Biosciences, San Jose, CA). Appropriate compensation was set by using MDA-MB-231 cells treated with the mitochondrial uncoupler FCCP (50 µM, 6 h) as positive control.…”

Data on cytotoxic activity of an Artemisia annua herbal preparation and validation of the quantification method for active ingredient analysis

“…Briefly, MDA-MB-231 cells were treated with Momundo (100 μg/ml), Momundo-ACN (30 μg/ml), paclitaxel 100 nM or 0.5% DMSO for control for 24 h. Then, cells were harvested by trypsinization and fixed with ice-cold ethanol (70%) overnight. DNA was stained with propidium iodide (43 μg/ml) in a buffer containing DNAse-free RNase A [3,4] and analyzed by flow cytometry (FACSVerse cytometer, Becton Dickinson, Heidelberg, Germany) and FlowJo software (FlowJo LLC, Ashland, OR).…”