Evaluating the Cytotoxic and Pro-Apoptotic Effects of Artemisinin on Human Breast Cancer Cell lines via MTT, Caspase Assays, and Gene Expression Studies

BORAH et al.: CYTOTOXIC AND APOPTOTIC EFFECTS OF ARTEMISININ IN CANCER CELLS

Abstract

Artemisinin, a sesquiterpene lactone derived from Artemisia annua, has demonstrated promising anti-cancer properties. This study aimed to evaluate the compound's pro-apoptotic and cytotoxic potential in MCF-7 human breast cancer cells. MCF-7 cells were treated with varying concentrations of Artemisinin (0.78–100 µg/mL), and apoptosis was assessed

by measuring caspase-3, -8, and -9 activities at 24, 48, and 72 hours post-treatment. Cell viability and cytotoxicity were quantified using the MTT assay, and IC₅₀ values were calculated accordingly. Gene expression of BAX and BCL-2 was analyzed via qPCR using the ΔΔCt method. Two-way ANOVA was performed to assess the combined effects of Artemisinin concentration and exposure time. A consistent dose- and time-dependent increase in caspase activity was noted, with Caspase-9 showing the most marked elevation at 72 hours, indicating strong activation of the intrinsic pathway. Artemisinin treatment significantly reduced MCF-7 cell viability, particularly at 24 and 48 hours. IC₅₀ values were
0.84 µg/mL (24 h), 1.33 µg/mL (48 h), and 2.82 µg/mL (72 h), demonstrating a time-dependent reduction in sensitivity. Two-way ANOVA showed significant effects of both Artemisinin concentration and exposure time on viability and cytotoxicity, with a notable interaction for cytotoxicity (p = 0.0268). BAX expression was upregulated, while BCL-2
was downregulated in a concentration and time dependent manner. Artemisinin induces significant cytotoxicity and apoptosis in MCF-7 breast cancer cells through caspase activation and modulation of BAX/BCL-2 expression, supporting its potential as a promising anti-cancer therapeutic agent.