Exploring the effects of Asacoumarin B, alone and in combination with irradiation, on cervical carcinoma cells

Introduction: High-risk human papillomavirus (HPV) infection is the primary cause of cervical carcinoma, with rising incidence of HPV-associated cases reported in countries including Iran. Radiotherapy remains a cornerstone for treating locally advanced cervical cancer, yet radioresistance often limits its efficacy. Asacoumarin B, a sesquiterpene coumarin from Ferula species, exhibits antiproliferative and pro-apoptotic activity across cancer models; this study investigates its effects, alone and combined with irradiation, on cervical carcinoma cells in vitro. Methods: HeLa cells, a human cervical carcinoma line, were seeded in ۹۶-well plates and treated with Asacoumarin B (۲۵, ۵۰, or ۱۰۰ μM) for ۲۴, ۷۲, or ۱۲۰ h at ۳۷°C (Memmert incubator). Cell viability was assessed via resazurin assay: resazurin solution was added, cells incubated for ۳ h, and absorbance measured at ۶۰۰ nm (Epoch plate reader). For combination therapy, cells pretreated with ۵۰ or ۱۰۰ μM Asacoumarin B for ۴۸ h were irradiated with X-rays (۴۰۰ or ۸۰۰ cGy; Elekta Compact™ linear accelerator, Crawley), followed by ۷۲ h recovery. Additionally, apoptosis was evaluated post-treatment (alone or combined) using FITC-annexin V/propidium iodide staining and flow cytometry (BD FACSCalibur). Results: Our findings indicated that viability of cells was ۹۱.۶%, ۹۵.۳% and ۸۵.۷% after ۲۴, ۷۲ and ۱۲۰ h treatment with the highest dose of Asacoumarin B (۱۰۰ μM), respectively. For combinatorial treatments with X-ray, ۴۸ h pretreatment with ۱۰۰ μM Asacoumarin B followed by ۴۰۰ and ۸۰۰ cGy irradiation decreased viability of cells to ۸۱.۳% and ۷۹.۸%, respectively. Additionally, flow cytometry analysis revealed ۶.۷% apoptosis in cells treated with ۱۰۰ μM Asacoumarin B for ۱۲۰ h, while this amount was ۱۳.۳% for cells treated with ۱۰۰ μM Asacoumarin B and ۸۰۰ cGy radiation. Conclusion: These results demonstrate that Asacoumarin B enhances irradiation efficacy in cervical carcinoma cells via reduced viability and increased apoptosis. Future studies with diverse cell lines and semisynthetic derivatives may improve therapeutic specificity.

کلیدواژه ها:

Asacoumarin B ، Cervical carcinoma ، Irradiation ، Viability assay ، Apoptosis