Modulating SASP-Driven Paracrine Effects to Overcome Therapy-Induced Senescence: Senolytic Role of Hesperidin and Quercetin in A549 Lung Cancer Cells

Purpose Therapy-induced senescence (TIS) is a double-edged outcome of chemotherapeutic stress, leading to durable cell cycle arrest and the secretion of pro-inflammatory SASP factors that may promote tumour progression and relapse. This study aimed to investigate the senolytic potential of quercetin (QCT) and hesperidin (HDN)—two flavonoids with known anticancer properties—in reversing 5-fluorouracil (5-FU)-induced senescence and SASP-mediated paracrine resistance in A549 lung adenocarcinoma cells. Methods A549 cells were treated with sublethal doses of 5-FU to induce senescence, confirmed by SA-β-gal staining and senescence marker expression (p16, p21). SASP-conditioned media (SASP-CM) were collected from senescent cultures and used to assess chemoresistance, migration, and gene expression profiles. The senolytic efficacy of QCT and HDN was evaluated using MTT assays, scratch assays, AO/EB staining, colony formation assays, X-Gal staining, and RT-PCR. Results Sublethal 5-FU induced robust senescence and increased resistance to subsequent 5-FU exposure in SASP-CM–treated A549 cells. Both QCT and HDN significantly reversed SASP-mediated chemoresistance and inhibited cell migration. HDN exhibited greater pro-apoptotic and senolytic activity than QCT, as evidenced by higher apoptosis rates, enhanced p53 expression, and reduced SASP marker expression. X-Gal and colony formation assays confirmed selective clearance of senescent cells. Conclusion This study demonstrates that HDN and QCT, particularly HDN, possess strong senolytic and SASP-suppressing effects, thereby restoring chemosensitivity in TIS-affected lung cancer cells. These findings support the use of flavonoid-based senotherapeutics as adjunct strategies to overcome therapy-induced resistance and tumour recurrence.