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外泌体转运的 circ_0081069 通过结合 miR-195-5p 诱导 SPIN1 的产生,从而抑制食管鳞癌细胞的放射敏感性。

Exosome-transported of circ_0081069 induces SPIN1 production by binding to miR-195-5p to inhibit radiosensitivity in esophageal squamous cell carcinoma.

机构信息

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.

出版信息

J Biochem Mol Toxicol. 2024 Feb;38(2):e23659. doi: 10.1002/jbt.23659.

Abstract

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.

摘要

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。

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