| Lin Chuanchuan,Yu Jiawu,Yang Zhenxing,et al.Mechanisms behind the influence of growth differentiation factor 15 on the radioresistance of osteosarcoma cells[J].Chinese Journal of Radiological Medicine and Protection,2026,46(6):564-572 |
| Mechanisms behind the influence of growth differentiation factor 15 on the radioresistance of osteosarcoma cells |
| Received:June 14, 2025 |
| DOI：10.3760/cma.j.cn112271-20250614-00201 |
| KeyWords:Osteosarcoma|Growth differentiation factor 15 (GDF15)|Radioresistance|Reactive oxygen species (ROS) |
| FundProject:重庆市自然科学基金面上项目(CSTB2022NSCQ-MSX0179, CSTB2023NSCQ-MSX0270);陆军军医大学第二附属医院青年博士人才孵化计划(2022YQB077, 2022YQB064);重庆市自然科学基金重点项目(cstc2020jcyj-zdxmX0013) |
| Author Name | Affiliation | E-mail | | Lin Chuanchuan | Department of Blood Transfusion, Laboratory Medicine Center, The Second Affiliated Hospital of AMU, Hematopoietic Acute Radiation Syndrome Medical and Pharmaceutical Basic Research Innovation Center, Ministry of Education of the People's Republic of China, Chongqing 400037, China | | | Yu Jiawu | Department of Blood Transfusion, Laboratory Medicine Center, The Second Affiliated Hospital of AMU, Hematopoietic Acute Radiation Syndrome Medical and Pharmaceutical Basic Research Innovation Center, Ministry of Education of the People's Republic of China, Chongqing 400037, China | | | Yang Zhenxing | Department of Blood Transfusion, Laboratory Medicine Center, The Second Affiliated Hospital of AMU, Hematopoietic Acute Radiation Syndrome Medical and Pharmaceutical Basic Research Innovation Center, Ministry of Education of the People's Republic of China, Chongqing 400037, China | | | Ran Qian | Department of Blood Transfusion, Laboratory Medicine Center, The Second Affiliated Hospital of AMU, Hematopoietic Acute Radiation Syndrome Medical and Pharmaceutical Basic Research Innovation Center, Ministry of Education of the People's Republic of China, Chongqing 400037, China | | | Li Zhongjun | Department of Blood Transfusion, Laboratory Medicine Center, The Second Affiliated Hospital of AMU, Hematopoietic Acute Radiation Syndrome Medical and Pharmaceutical Basic Research Innovation Center, Ministry of Education of the People's Republic of China, Chongqing 400037, China | | | Xiang Yang | Department of Blood Transfusion, Laboratory Medicine Center, The Second Affiliated Hospital of AMU, Hematopoietic Acute Radiation Syndrome Medical and Pharmaceutical Basic Research Innovation Center, Ministry of Education of the People's Republic of China, Chongqing 400037, China | xiangy105@163.com |
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| Abstract:: |
| Objective To explore the effects of the expression level of growth differentiation factor 15 (GDF15) on the radiosensitivity of osteosarcoma and the mechanisms behind the effects. Methods The cancerous and paracancerous tissues of osteosarcoma were collected, and the expression levels of GDF15 in these tissues were detected using immunohistochemistry and Western blot. Human osteosarcoma cell lines U2OS and Saos2 were randomly divided into four groups: negative control, GDF15 knockdown (i.e., siGDF15), irradiation control, and combined treatment groups. Among these, the irradiation control group received 9 Gy of γ-ray irradiation. The effects of irradiation on GDF15 expression levels in U2OS and Saos2 were then detected using quantitative polymerase chain reaction (qPCR) and Western blot. Using method including flow cytometry, immunofluorescence assay, and Western blot, the apoptosis rates and the expression levels of apoptosis-related proteins in various groups were assessed following 9 Gy of irradiation. Subsequently, the radiation stress genes and signaling pathways associated with GDF15 were selected through transcriptome sequencing analysis. Additionally, the effects of siGDF15 on key molecules after irradiation were verified using qPCR, Western blot, and immunofluorescence assay. Results The four groups of clinical samples of paired cancerous tissues exhibited significantly higher mRNA and protein expression levels of GDF15 than the paracancerous tissues (tmRNA = 5.33-16.32, P < 0.05; tprotein = 5.83-13.49, P < 0.05). Furthermore, irradiation could upregulate the mRNA expression levels of GDF15 in U2OS and Saos2 over time of continued culture (tU2OS= 12.9-54.71, P < 0.05; tSaos2= 5.46-37.69, P < 0.05). Compared to the irradiation control group, the combined treatment group showed significantly upregulated apoptosis (t = 10.93, P < 0.05), increased proportions of cells in the G2/M phases (t = 15.63, P < 0.05), and significantly increased number of γ-H2AX foci at 24 h after irradiation (t = 15.54, P < 0.05). Transcriptome sequencing analysis suggested the enrichment in reactive oxygen species (ROS)-related signaling pathways, which was consistent with the qPCR validation result. The analytical result also indicated that ROS-related genes, including ACSL4, SLC7A11, GPX4, and SLC3A2, were significantly altered by siGDF15. Compared to the irradiation control group, the combined treatment group showed elevated ROS level and malondialdehyde (MDA) content, as well as reduced mitochondrial membrane potential (MMP) and glutathione (GSH) level, after radiation. Besides, the addition of ponsegromab, an exogenous GDF15 monoclonal antibody, decreased the protein level of NRF2 and increased the number of γ-H2AX foci and the apoptosis rate after radiation. In contrast, the addition of recombinant GDF15 protein increased the protein level of NRF2, decreased the number of γ-H2AX foci, and significantly reduced the apoptosis rate after radiation (t = 21.32, P < 0.05). Conclusions GDF15 is highly expressed in the cancerous tissues of osteosarcoma. Following radiation, GDF15 manifests an upregulated expression level and participates in oxidative stress response through NRF2- and ROS-related genes. Knockdown of GDF15 can enhance the radiosensitivity of osteosarcoma cells. |
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