Abstract

Introduction: Breast cancer remains a significant challenge due to disease recurrence and metastasis, often attributed to the persistence of treatment-resistant breast cancer stem cells (BCSCs). Citral, a compound derived from lemongrass essential oil, has demonstrated cytotoxic effects on various cancers, including breast cancer. This study investigates the anticancer mechanisms of Citral in BCSC-enriched 3D cultured spheroids and evaluates its therapeutic potential for estrogen receptor-positive breast cancer.

Methods: Using flow cytometry, the CD44⁺CD24^- population was analyzed, and real-time PCR was employed to measure the expression of ALDH isoforms and pluripotency genes. Additionally, the Nanostring nCounter® PanCancer Pathway Panel was utilized to identify gene expression changes in cancer-related pathways.

Results: Citral treatment significantly reduced spheroid size and the CD44⁺CD24^- stem-like cell population, accompanied by downregulation of ALDH isoforms and pluripotency genes. Gene expression analysis revealed Citral's modulation of key pathways, including PI3K/Akt signaling, cell cycle control, DNA damage response, and apoptosis.

Conclusion: These findings underscore Citral's potential as a promising anticancer agent, particularly for targeting estrogen receptor-positive breast cancer cells and BCSCs. Further preclinical and clinical studies are warranted to explore its therapeutic applications.