In Vitro Antioxidant and p53-Targeted Anticancer Activity of Aerva sanguinolenta (L.) Blume Across Different Human Cancer Cell Lines and Ehrlich Ascites Carcinoma Model

Abstract

This study investigated the antioxidant, cytotoxic, and antitumor activities of Aerva sanguinolenta (L.) Blume extracts using in vitro, in vivo, and in silico approaches to explore its potential as a natural anticancer agent. Crude methanolic extract (CME) and its petroleum ether (PEF), chloroform (CHF), ethyl acetate (EAF), and aqueous (AQF) fractions were analyzed. Antioxidant activities were evaluated using phosphomolybdenum, ferric-reducing power, DPPH, hydroxyl radical scavenging, and lipid peroxidation assays. Cytotoxicity was assessed using MTT assays against HeLa (cervical), H-2228 and H-3122 (lung), and HEK-293 (kidney epithelial) cell lines. In vivo anticancer evaluation was performed in Ehrlich ascites carcinoma (EAC)–bearing mice. GC–MS identified chemical constituents, followed by p53-targeted molecular docking and ADMET predictions. CHF showed the highest phenolic and flavonoid contents, correlating with superior antioxidant activity. MTT assays revealed strong dose-dependent antiproliferative effects of CME, CHF, and EAF against HeLa cells (IC50 = 17–21 μg/mL), comparable to vincristine sulfate (VS) and 5-fluorouracil (5-FU). CHF also exhibited potent cytotoxicity against H-2228, H-3122, and HEK-293 cells, demonstrating broad-spectrum anticancer activity. In the EAC mouse model, CHF (10 mg/kg) inhibited tumor growth by 77.2%, approaching the efficacy of bleomycin. GC–MS analysis identified 17 compounds, among which 2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl-, (R)- showed the strongest p53-binding affinity. ADMET predictions indicated favorable pharmacokinetic and toxicity profiles for key candidates. A. sanguinolenta, particularly its chloroform fraction, shows strong antioxidant and p53-mediated anticancer potential, supporting its promise as a natural source for developing novel anticancer agents.