| 杜梦洁,于祖祥,马合丽亚·阿地力,等.活性氧物种调节因子1对人皮肤黑色素瘤细胞的放射增敏作用研究[J].中华放射医学与防护杂志,2026,46(8):731-739.Du Mengjie,Yu Zuxiang,Adili Maheliya,et al.Radiosensitization effects of reactive oxygen species modulator 1 on human cutaneous melanoma cells[J].Chin J Radiol Med Prot,2026,46(8):731-739 |
| 活性氧物种调节因子1对人皮肤黑色素瘤细胞的放射增敏作用研究 |
| Radiosensitization effects of reactive oxygen species modulator 1 on human cutaneous melanoma cells |
| 投稿时间：2025-10-03 |
| DOI：10.3760/cma.j.cn112271-20251003-00353 |
| 中文关键词: 黑色素瘤|活性氧物种调节因子1|放射敏感性|细胞凋亡|活性氧 |
| 英文关键词:Melanoma|Reactive oxygen species modulator 1|Radiosensitivity|Apoptosis|Reactive oxygen species |
| 基金项目:国家自然科学基金(82473574,82404200,U25A20151);四川省自然科学基金青年科学基金(25QNJJ4600) |
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| 中文摘要: |
| 目的 研究活性氧物种调节因子1(ROMO1)对黑色素瘤细胞放射敏感性的影响,并初步探讨其机制,为黑色素瘤放射增敏剂的研发提供实验依据。方法 以人皮肤黑色素瘤细胞A375为研究对象,按照射与病毒处理方式分为4组:空载病毒组(Ad-NC)、ROMO1过表达组(Ad-ROMO1)、空载+照射组(Ad-NC+IR)和ROMO1过表达+照射组(Ad-ROMO1+IR)。采用10 Gy X射线进行照射。通过CCK-8法、乳酸脱氢酶(LDH)释放实验、平板克隆形成实验、细胞划痕实验以及流式细胞术等手段,检测ROMO1过表达对照后A375细胞存活、增殖、迁移、凋亡、活性氧(ROS)生成及线粒体膜电位的影响。利用Western blot检测与细胞死亡相关的蛋白表达水平,RNA-seq筛选差异表达基因进行通路富集分析。结果 给予10 Gy X射线照射,相较Ad-NC细胞,ROMO1过表达会抑制A375细胞活力(t = 7.62、7.25,P <0.05),增加LDH释放(t = 4.92、3.77,P <0.05),抑制细胞克隆形成(t = 2.77、8.07、9.90,P <0.05)及细胞迁移(t = 8.19,P <0.05),同时促进细胞凋亡(t = 4.32、26.01,P <0.05),增加ROS水平(t = 25.12,P <0.05),降低线粒体膜电位(t = 13.77、8.98,P <0.05),呈现明显的放射增敏效应。Western blot显示,受照后Ad-ROMO1细胞较Ad-NC cleaved-caspase 3、cleaved-PARP表达上调,BAX/Bcl-2比值升高。RNA-seq表明,10 Gy照射后,Ad-ROMO1与Ad-NC相比共鉴定出97个差异表达基因(58个上调和39个下调);京都基因与基因组百科全书(KEGG)提示上调差异基因主要富集于神经活性配体信号、系统性红斑狼疮、细胞凋亡、细胞周期及JAK-STAT/Wnt信号通路等,下调差异基因则主要富集于病毒感染通路。结论 过表达ROMO1使得人黑色素瘤细胞的放射敏感性增强,其作用可能与调控ROS生成、促进凋亡及影响免疫炎症相关信号通路有关。 |
| 英文摘要: |
| Objective To investigate the effects of reactive oxygen species modulator 1 (ROMO1) on the radiosensitivity of melanoma cells and to preliminarily explore the mechanisms underlying the effects, in order to provide experimental evidence for the research and development of radiosensitizers for melanoma treatment. Methods Human cutaneous melanoma cells A375 were investigated in this study. Based on irradiation and adenovirus treatment method, the cells were divided into four groups, namely the adenovirus negative control group (Ad-NC), the ROMO1 overexpression group (Ad-ROMO1), the adenovirus negative + irradiation group (Ad-NC + IR), and the ROMO1 overexpression + irradiation group (Ad-ROMO1 + IR), with 10 Gy X-ray irradiation adopted. Using the cell counting Kit-8 (CCK-8) assay, lactate dehydrogenase (LDH) release assay, plate colony formation assay, wound healing assay, and flow cytometry, the effects of ROMO1 overexpression on the survival, proliferation, migration, apoptosis, reactive oxygen species (ROS) generation, and the mitochondrial membrane potential of the A375 cells were assessed. The expression levels of cell death related proteins were detected using Western blot analysis. Furthermore, differentially expressed genes (DEGs) were selected using the RNA sequencing (RNA-seq) method, followed by pathway enrichment analysis. Results Compared with the Ad-N + IR group, the Ad-ROMO1 + IR group exhibited suppressed viability of the A375 cells (t = 7.62, 7.25, P < 0.05), increased LDH release (t = 4.92, 3.77, P < 0.05), inhibited colony formation (t = 2.77, 8.07, 9.90, P < 0.05), and decreased cell migration (t = 8.19, P<0.05), as well as promoted apoptosis (t = 4.32, 26.01, P < 0.05), elevated ROS levels (t = 25.12, P < 0.05), and decreased mitochondrial membrane potential (t = 13.77, 8.98, P < 0.05). These results demonstrated distinct radiosensitization effects of ROMO1 overexpression. Western blot analysis indicated that after 10 Gy X-ray irradiation, the Ad-ROMO1 group showed upregulated expression of cleaved-caspase 3 and cleaved-PARP and an increased BAX/Bcl-2 ratio compared to the Ad-NC group. The analysis results derived using the RNA-seq method indicated that after irradiation, the Ad-ROMO1 group exhibited a total of 97 DEGs (58 upregulated and 39 downregulated genes) compared to the Ad-NC group. KEGG pathway enrichment analysis indicated that the upregulated DEGs were mainly enriched in signaling pathways involved in neuroactive ligand signaling, systemic lupus erythematosus (SLE), apoptosis, and cell cycle, as well as the JAK-STAT/Wnt signaling pathway. In contrast, the downregulated DEGs were primarily enriched in adenovirus infection-related pathways.Conclusions ROMO1 overexpression can enhance the radiosensitivity of human melanoma cells, potentially functioning by regulating ROS generation, promoting apoptosis, and modulating immune and inflammation-related signaling pathways. |
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