Pszczółkowska-Kępa Beata, Olejarz Wioletta, Głuszko Alicja, Wałpuski Grzegorz, Lorenc Tomasz, Brzozowska Beata
Biomedical Physics Division, Faculty of Physics, University of Warsaw, Warsaw, Poland.
Department of Biochemistry and Pharmacogenomics, Faculty of Pharmacy, Medical University of Warsaw, Warsaw, Poland.
PLoS One. 2025 Aug 28;20(8):e0330501. doi: 10.1371/journal.pone.0330501. eCollection 2025.
Exosomes are involved in intracellular communication and mediate the radiation-induced bystander effect (RIBE). We assessed the ability of exosomes to modify the radiation response of PC3 and DU145 prostate cancer cells exposed to X-rays. Irradiated cells were analyzed using clonogenic survival and apoptosis assays, while exosome-stimulated cells were evaluated for γH2AX immunostaining, immunoblotting, and apoptosis. Exosomes were isolated via size exclusion chromatography (SEC), characterized by nanoparticle tracking analysis (NTA) and immunoblotting, and ranged from 130 to 137 nm, containing CD63 and CD81. We found that exposure to ionizing radiation (IR) resulted in increased apoptosis and necrosis. To assess exosomes impact on radiation response, exosomes were transferred to non-irradiated and irradiated recipient cells. Non-irradiated PC3 cells stimulated by exosomes released from irradiated PC3 and DU145 cells showed more apoptosis and necrosis than those stimulated by exosomes released from non-irradiated cells. Non-irradiated PC3 cells co-incubated with exosomes from irradiated PC3 and DU145 cells exhibited more γH2AX foci than non-irradiated PC3 cells. Our results confirmed that DU145 cells are more radioresistant than PC3 cells and exosomes isolated from these cells may contribute to radiation resistance in prostate cancer. Thus, studying exosome functions, particularly in radiation resistance, is crucial for understanding carcinogenesis and optimizing radiotherapeutic methods.
外泌体参与细胞内通讯并介导辐射诱导的旁观者效应（RIBE）。我们评估了外泌体对暴露于X射线的PC3和DU145前列腺癌细胞辐射反应的影响。使用克隆形成存活和凋亡分析来分析受辐照的细胞，而对外泌体刺激的细胞进行γH2AX免疫染色、免疫印迹和凋亡评估。通过尺寸排阻色谱法（SEC）分离外泌体，通过纳米颗粒跟踪分析（NTA）和免疫印迹进行表征，其大小范围为130至137nm，含有CD63和CD81。我们发现暴露于电离辐射（IR）会导致凋亡和坏死增加。为了评估外泌体对辐射反应的影响，将外泌体转移至未受辐照和受辐照的受体细胞。由辐照后的PC3和DU145细胞释放的外泌体刺激的未受辐照的PC3细胞比由未受辐照细胞释放的外泌体刺激的细胞表现出更多的凋亡和坏死。与来自辐照后的PC3和DU145细胞的外泌体共同孵育的未受辐照的PC3细胞比未受辐照的PC3细胞表现出更多的γH2AX焦点。我们的结果证实，DU145细胞比PC3细胞具有更高的放射抗性，从这些细胞中分离的外泌体可能有助于前列腺癌中的放射抗性。因此，研究外泌体功能，特别是在放射抗性方面，对于理解致癌作用和优化放射治疗方法至关重要。
