Dilay Karademir¹ , Ay?egül Öztürk², Bilal ?ahin³, Mustafa Özkaraca⁴

Abstract

Purpose: To investigate the roles that autophagy, oxidative DNA damage, and proapoptotic and anti-apoptotic proteins play in astaxanthin's cytotoxic action on HeLa cells. Methods: HeLa cells were treated with varying concentrations of astaxanthin (2.5 - 40 μM). Untreated cells served as the negative control. Cell viability was assessed using XTT assay to determine the cytotoxic effects of the compound. The levels of B-cell lymphoma 2 (BCL-2), Bcl-2-associated X protein (Bax), caspase 3, poly (ADP-ribose) polymerase (PARP), and 8-hydroxy-2'-deoxyguanosine (8-oxo-dG) were measured by ELISA using cell lysates collected after a 24-hour incubation with astaxanthin at IC50 concentration. Furthermore, immunofluorescence staining was performed to evaluate the expression of 8-oxo-dG and Microtubule-associated protein 1 light chain 3 beta (LC3β) in HeLa cells. Results: Astaxanthin exhibited a concentration-dependent cytotoxic effect on HeLa cells, with an IC50 value of 18.47 μM after 24 hours. Treatment with IC50 dose significantly increased the expression of apoptotic proteins (cleaved caspase 3, Bax, cleaved PARP; p < 0.001) while leaving BCL-2 expression unchanged (p > 0.05). Furthermore, the use of this medicine resulted in an enhancement in the levels of 8-oxo-dG (p < 0.001), a well-recognized indicator that signifies oxidative harm to DNA. Additionally, there was an observed rise in LC3β levels (p < 0.05), an indicator closely linked to the process of autophagy. Conclusion: Astaxanthin's cytotoxic activity against cervical cancer may be attributed to its capacity to induce both apoptotic and autophagic cell death pathways. This finding emphasizes the positive potential of astaxanthin as a cervical cancer therapy option.