细胞骨架调节剂对脊索瘤细胞生物学行为及放射敏感性的影响  

CYTOR regulates the biological behavior and radiosensitivity of human chordoma cell lines

作　　者：姚杰 秦利[2] 吴学建[2] Yao Jie;Qin Li;Wu Xuejian(Spinal surgery,Luoyang Orthopedic-Traumatological Hospital of Henan Province(Henan Provincial Orthopaedic Hospital),Zhengzhou 450000,China)

机构地区：[1]河南省洛阳正骨医院(河南省骨科医院)脊柱外科,郑州450000 [2]郑州大学第一附属医院骨科,450052

出　　处：《中华骨科杂志》2020年第13期856-863,共8页Chinese Journal of Orthopaedics

摘　　要：目的通过实验研究探讨细胞骨架调节剂(lncRNA cytoskeleton regulator,CYTOR)对脊索瘤CM-319和U-CH1细胞生物学行为,包括增殖、迁移和侵袭,及放射敏感性的影响及潜在的分子机制。方法脊索瘤组织和髓核组织中CYTOR和miR-24-3p的表达采用qRT-PCR检测。将人脊索瘤细胞株CM-319和U-CH1分为si-NC组(转染si-NC)、si-CYTOR组(转染si-CYTOR)、miR-NC组(转染miR-NC)、miR-24-3p组(转染miR-24-3p)、si-CYTOR+anti-miR-NC组(共转染si-CY-TOR和anti-miR-NC)、si-CYTOR+anti-miR-24-3p组(共转染si-CYTOR和anti-miR-24-3p)。Western blot检测增殖、迁移侵袭蛋白表达水平,MTT法测定CM-319和U-CH1细胞增殖活性,Transwell实验检测细胞迁移和侵袭能力,克隆形成实验检测不同剂量放射处理后细胞存活分数,双荧光素酶报告系统验证CYTOR和miR-24-3p的调控关系。结果检测20个脊索瘤组织和20个髓核组织样本,结果发现,lncRNA CYTOR在脊索瘤组织中表达量显著高于髓核组织(t=19.837,P<0.05),miR-24-3p在脊索瘤组织中表达量显著低于髓核组织(t=22.061,P<0.05);抑制CYTOR表达和过表达miR-24-3p均可抑制CM-319和U-CH1细胞增殖、迁移、侵袭,增强细胞放射敏感性;双荧光素酶报告实验结果表明,lncRNACYTOR靶向负调控miR-24-3p的表达;抑制miR-24-3p逆转了抑制lncRNA CYTOR对CM-319和U-CH1细胞增殖、迁移、侵袭和放射敏感性的作用。结论lncRNA CYTOR通过靶向调控miR-24-3p表达抑制CM-319和U-CH1细胞的增殖、迁移和侵袭,增强细胞放射敏感性。lncRNA CYTOR是脊索瘤潜在的分子治疗靶点。Objective To investigate the effects of lncRNA cytoskeleton regulator (CYTOR) on proliferation, migration, invasion and radiosensitivity in chordoma CM-319 and U-CH1cells and uncover the underlying mechanism.Methods The expression of CYTOR and miR-24-3p in chordoma and nucleus pulposus tissues were detected by qRT-PCR. Human chordoma cell lines CM-319 and U-CH1 were divided into si-NC group (transfected si-NC), si-CYTOR group (transfected si-CYTOR), miR-NC group (transfected miR-NC), MiR-24-3p group (transfected miR-24-3p), si-CYTOR+anti-miR-NC group (co-transfected with si-CYTOR and anti-miR-NC), si-CYTOR+anti-miR- Group 24-3p (co-transfected with si-CYTOR and anti-miR-24-3p). Western blot was applied to detect the expression levelsof proteinsrelatedtoproliferation, migration and invasion. MTT was used to measure the proliferation of CM-319 and U-CH1 cells. Transwell assay was implemented to detect the migration and invasion ability of cells. Colony formation assay was used to detect the survival fraction of CM-319 and U-CH1 cells after different doses of radiation treatment. The relationship between CYTOR and miR-24-3p was verified by dual-luciferase reporter assay system.Results The detection of 20 samples of chordoma tissues and 20 samples of nucleus pulposus tissuesrevealed that the expression level of CYTOR in chordoma tissues was significantly higher than that in nucleus pulposus tissues (t=19.837, P<0.05), and the expression level of miR-24-3p in chordoma tissues was significantly lower than that in nucleus pulposus tissues (t=22.061, P<0.05). Either inhibiting CYTOR or overexpression of miR-24-3p inhibited the proliferation, migration and invasion of CM-319 and U-CH1 cells, enhanced the radiosensitivity of CM-319 and U-CH1 cells. The result of dual-luciferase reporter assay system suggested that CYTOR negatively regulated the expression of miR-24-3p. Inhibition of miR-24-3p reversed the effects of inhibiting CYTOR on the proliferation, migration, invasion and radiosensitivity of CM-319 and U-CH1 cells.Co

关 键 词：脊索瘤 细胞骨架 细胞增殖 细胞迁移分析 肿瘤侵润 辐射耐受性 

分 类 号：R738.1[医药卫生—肿瘤]