Department of Radiotherapy, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Department of Radiotherapy, The Second Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
J Biochem Mol Toxicol. 2024 Feb;38(2):e23659. doi: 10.1002/jbt.23659.
Circ_0081069 plays a key role in tumor growth; however, its effect on radiosensitivity in esophageal squamous cell carcinoma (ESCC) remains unknown. The study is performed to reveal the association of circ_0081069 expression and radiosensitivity in ESCC and the underlying mechanism. Circ_0081069, miR-195-5p, and spindlin 1 (SPIN1) RNA expression were detected by quantitative real-time polymerase chain reaction. Protein expression was checked by Western blot analysis or immunohistochemistry assay. Cell viability, proliferation, cell apoptosis, migration, and invasion were investigated by cell counting kit-8, 5-Ethynyl-29-deoxyuridine, flow cytometry analysis, scratch test, and transwell assays, respectively. The sensitivity of ESCC cells to radiation was investigated by cell colony formation assay. The interactions among circ_0081069, miR-195-5p, and SPIN1 were identified by dual-luciferase reporter assay and RNA Immunoprecipitation assay. Xenograft mouse model assay was performed to determine the effect of circ_0007841 on radiosensitivity in vivo. Circ_0081069 and SPIN1 expression were upregulated, whereas miR-195-5p was downregulated in ESCC tissues, ESCC cells, and radiation-stimulated ESCC cells. Circ_0081069 silencing inhibited ESCC cell proliferation, invasion, and migration but improved cell apoptosis. In addition, circ_0081069 knockdown enhanced ESCC cell radiosensitivity in vitro and in vivo. Circ_0081069 bound to miR-195-5p and regulated radiosensitivity by binding to miR-195-5p in ESCC cells. Moreover, SPIN1, a target of miR-195-5p, rescued miR-195-5p-mediated effects in ESCC cells. Circ_0081069 was secreted from ESCC cells by being packaged into exosomes. Further, circ_0081069-Exo inhibited radiosensitivity in ESCC cells. Exosome-mediated transfer of circ_0081069 induced SPIN1 production by binding to miR-195-5p, further inhibiting radiosensitivity in ESCC.
circ_0081069 在肿瘤生长中发挥关键作用；然而，其对食管鳞状细胞癌 (ESCC) 放射敏感性的影响尚不清楚。本研究旨在揭示 circ_0081069 表达与 ESCC 放射敏感性的关联及其潜在机制。通过实时定量聚合酶链反应检测 circ_0081069、miR-195-5p 和旋毛虫 1 (SPIN1) RNA 的表达。通过 Western blot 分析或免疫组织化学检测蛋白质表达。通过细胞计数试剂盒-8、5-乙炔基-29-脱氧尿苷、流式细胞术分析、划痕试验和 Transwell 试验分别研究细胞活力、增殖、细胞凋亡、迁移和侵袭。通过细胞集落形成试验研究 ESCC 细胞对辐射的敏感性。通过双荧光素酶报告基因试验和 RNA 免疫沉淀试验鉴定 circ_0081069、miR-195-5p 和 SPIN1 之间的相互作用。通过异种移植小鼠模型试验确定 circ_0007841 在体内对放射敏感性的影响。circ_0081069 和 SPIN1 的表达在 ESCC 组织、ESCC 细胞和辐射刺激的 ESCC 细胞中上调，而 miR-195-5p 的表达下调。circ_0081069 沉默抑制 ESCC 细胞增殖、侵袭和迁移，但促进细胞凋亡。此外，circ_0081069 敲低增强了 ESCC 细胞在体外和体内的放射敏感性。circ_0081069 与 miR-195-5p 结合，并通过结合 ESCC 细胞中的 miR-195-5p 调节放射敏感性。此外，miR-195-5p 的靶标 SPIN1 挽救了 miR-195-5p 在 ESCC 细胞中的作用。circ_0081069 被包裹在 ESCC 细胞的外泌体中从 ESCC 细胞中分泌出来。此外，circ_0081069-Exo 通过与 miR-195-5p 结合抑制 ESCC 细胞的放射敏感性。外泌体介导的 circ_0081069 转移通过与 miR-195-5p 结合诱导 SPIN1 产生，从而进一步抑制 ESCC 的放射敏感性。
