Flavonoid-Enriched Extract of Carissa carandas Linn Triggers Apoptosis and Cell Cycle Arrest in A549 Lung Cancer Cells: An Integrated In-Vitro and In-silico Approach Targeting CDK2 and Bcl-2
TL;DR
The findings suggest that EECC exerts anticancer effects against A549 lung carcinoma cells primarily through ROS-mediated apoptosis and cell cycle arrest.
Abstract
Carissa carandas Linn., a flavonoid-rich medicinal plant, has been reported to exhibit diverse pharmacological activities, including anticancer potential. Given that lung cancer remains one of the leading causes of cancer-related mortality worldwide and current chemotherapeutic regimens are often limited by toxicity and resistance, exploring plant-based alternatives offers a promising approach for developing safer and more effective anticancer therapies. This study was anticipated to evaluate the anticancer activity of ethanol-based extract of Carissa carandas leaves (EECC) on A549 human lung carcinoma cells, focusing on cytotoxicity, apoptosis induction, ROS generation, cell cycle arrest, and molecular interactions of key phytoconstituents with cancer-related targets, and to predict their pharmacokinetic and drug-likeness properties through in-silico methods. Phytochemical screening and antioxidant assays were performed initially. Cytotoxicity was assessed using the MTT assay, and apoptosis was confirmed by DAPI staining and Western blot analysis of apoptosis-related proteins (caspase-3, Bax, and Bcl-2). In-silico molecular docking was conducted using SwissDock for seven phytochemicals against cancer-related targets CDK2 and Bcl-2. SwissADME was employed to predict pharmacokinetic properties, including oral bioavailability, using bioavailability radar and BOILED-Egg models. EECC exhibited dose-dependent cytotoxicity with an IC₅₀ of 70.00 ± 1.50 µg/mL. Significant induction of apoptosis was observed, associated with elevated ROS levels (p < 0.01) and G0/G1 cell cycle arrest. Western blot analysis revealed upregulation of Bax and caspase-3 with concomitant downregulation of Bcl-2. Molecular docking identified Carindone and Quercetin as potent bioactive constituents showing strong binding affinities toward CDK2 and Bcl-2. SwissADME analysis confirmed favourable pharmacokinetic and oral bioavailability profiles of these compounds. EECC demonstrated dose-dependent cytotoxicity (IC50 of 87.96 ± 0.64 μg/mL). Apoptotic markers showed increased Bax and caspase-3 levels and decreased expression of Bcl-2. Apoptosis was significantly induced at increasing concentrations, correlating with elevated ROS levels (p < 0.01) and G0/G1 phase arrest. Docking analysis identified Carindone and Quercetin as key bioactive compounds, with strong binding affinities to CDK2 and Bcl-2. ADME analysis revealed favourable drug-likeness and oral bioavailability profiles for these compounds. The findings suggest that EECC exerts anticancer effects against A549 lung carcinoma cells primarily through ROS-mediated apoptosis and cell cycle arrest. The molecular docking outcomes strengthen the experimental observations by highlighting Carindone and Quercetin as potential modulators of cancer-associated proteins. The consistency between in-vitro and in-silico analyses underscores the therapeutic potential of EECC and its phytoconstituents as promising anticancer leads. EECC suppresses A549 lung cancer cell growth via ROS-mediated apoptosis and cell cycle arrest. Computational studies support these findings, identifying Carindone and Quercetin as promising lead compounds with desirable pharmacokinetic properties. These results warrant further in vivo and clinical analysis to explore the therapeutic potential of EECC in lung cancer treatment.