miR-34a降低人骨肉瘤肿瘤干细胞的自我更新能力并抑制其成瘤和迁移

发布时间：2018-04-25 15:59

本文选题：骨肉瘤 + 肿瘤干细胞　；参考：《细胞与分子免疫学杂志》2017年06期

【摘要】：目的确定从人骨肉瘤U2OS细胞群中分离培养的肿瘤干细胞中微小RNA-34a(miR-34a)的水平变化,并确定其在肿瘤干细胞成瘤性、干性维持中的作用。方法采用无血清培养法从U2OS细胞系中富集具有自我更新能力的肿瘤干细胞亚群,实时定量PCR检测miR-34a的水平;转染miR-34a模拟物(miR-34a mimics)或SOX2短发夹RNA(shRNA)后,Western blot法检测miR-34a靶蛋白性别决定区Y盒2(SOX2)蛋白水平,肿瘤干细胞微球形成实验检测自我更新能力,软琼脂成瘤实验检测miR-34a对肿瘤干细胞体外成瘤能力的影响,TranswellTM迁移实验检测miR-34a对肿瘤干细胞迁移能力的影响。结果骨肉瘤肿瘤干细胞中miR-34a水平降低;转染miR-34a mimics和转染SOX2 shRNA(sh SOX2)均降低SOX2蛋白水平,并降低肿瘤干细胞的自我更新能力、体外成瘤活性及迁移能力。结论骨肉瘤肿瘤干细胞中的miR-34a具有降低干细胞自我更新能力、成瘤性和迁移能力的作用。
[Abstract]:Objective to determine the changes in the level of small RNA-34a (miR-34a) in the human osteosarcoma U2OS cell group, and to determine its role in the tumorigenicity and dry maintenance of the tumor stem cells. Methods the serum-free culture method was used to enrich the subgroup of tumor stem cells with self renewal ability from the U2OS cell line, and the quantitative PCR was quantified in real time. The level of miR-34a was detected; after transfection of miR-34a analogue (miR-34a mimics) or SOX2 short hairpin RNA (shRNA), Western blot method was used to detect Y box 2 (SOX2) protein level of miR-34a target protein, and the ability of tumor stem cell microspheres to test self renewal and the ability of tumor stem cells to be tumorigenic in vitro by soft agar test Influence, TranswellTM migration test detected the effect of miR-34a on the migration ability of tumor stem cells. Results the level of miR-34a in osteosarcoma stem cells decreased; miR-34a mimics transfected and transfected SOX2 shRNA (SH SOX2) decreased the level of SOX2 protein, and reduced the self-renewal ability of tumor stem cells, the tumor activity and migration ability in vitro. MiR-34a in sarcoma tumor stem cells can reduce the self-renewal ability, tumorigenicity and migration ability of stem cells.

【作者单位】：重庆三峡医药高等专科学校病原生物学与免疫学教研室;重庆三峡医药高等专科学校基础医学部病理学教研室;重庆三峡医药高等专科学校临床医学系;成都中医药大学附属医院;
【基金】：重庆三峡医药高等专科学校校级苗圃工程(2016mpxz19) 重庆市卫计委中医项目(zy201602100) 四川省科技支撑项目(2016FZ0093)
【分类号】：R738

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