代 国,刘盖为,孙祥然,胡庆柱,郑 迪,杨 俭,郭卫春.MiR-335通过作用于POU5F1负性调节骨肉瘤干细胞特性的实验研究[J].,2017,17(34):6607-6612 |
MiR-335通过作用于POU5F1负性调节骨肉瘤干细胞特性的实验研究 |
MiR-335 Negatively Regulates Osteosarcoma Stem Cell-like Properties by Targeting POU5F1 |
投稿时间:2017-05-30 修订日期:2017-06-25 |
DOI:10.13241/j.cnki.pmb.2017.34.002 |
中文关键词: 骨肉瘤;干细胞; miRNA-335 POU5F1 |
英文关键词: Osteosarcoma Cancer stem cell miR-335 POU5F1 |
基金项目:国家自然科学基金青年基金项目(81502575);中央高校基本科研业务费专项基金(2042015kf0069, 2042017kf0163) |
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中文摘要: |
摘要 目的:研究表明骨肉瘤干细胞与骨肉瘤的发生发展及耐药密切相关。本文探讨miR-335在骨肉瘤干细胞中所起的作用。方法:利用不同方法筛选出骨肉瘤干细胞,检测miR-335的表达水平。利用分子探针技术将骨肉瘤细胞分选为miR-335高表达组和低表达组,检测各组中骨肉瘤细胞的干细胞样特性。利用转染技术检测miR-335对骨肉瘤干细胞样特性的影响。利用荧光素酶报告基因系统,验证miR-335下游靶基因。最后,利用裸鼠移植瘤动物模型,检测pre-miR-335联合化疗药阿霉素对骨肉瘤的在体抑制效果。结果:miR-335在骨肉瘤干细胞中表达量明显减少。miR-335高表达细胞的干细胞特性明显降低。转染实验验证了上述结果。荧光素酶报告试验显示miR-335在转录后水平上负性调节POU5F1基因。动物实验表明pre-miR-335 联合阿霉素在抑制骨肉瘤上明显优于单一阿霉素的抑制效果。结论:miR-335通过下调 POU5F1负性调节骨肉瘤干细胞特性。miR-335 联合传统化疗药可协同抑制骨肉瘤。 |
英文摘要: |
ABSTRACT Objective: Studies have shown that osteosarcoma stem cells are highly correlated to the occurrence, development and chemoresistance of osteosarcoma. Thus, we investigate the role of miR-335 in osteosarcoma stem cells. Methods: The osteosarcoma stem cells were screened out by different methods, and the expression level of miR-335 was detected. The osteosarcoma cells were sorted into miR-335 high expression group and miR-335 low expression group by molecular probe technique, and the stem cell-like characteristics of osteosarcoma cells in each group were detected. The transfection technique was used to detect the effect of miR-335 on the characteristics of osteosarcoma stem cells. Luciferase reporter gene system was used to verify target genes downstream of miR-335. Finally, the nude mice xenograft model was used to detect the inhibitory effect of pre-miR-335 combined with chemotherapeutic agent doxorubicin on osteosarcoma. Results: The expression of miR-335 in osteosarcoma stem cells was significantly decreased. The stem cell characteristics of miR-335 overexpressing cells were significantly decreased. Transfection experiments confirmed the above results. Luciferase reporter test showed that miR-335 negatively regulates POU5F1 gene at post transcriptional level. Animal experiments showed that pre-miR-335 combined with doxorubicin significantly inhibited the inhibitory effect of single agent on osteosarcoma. Conclusion: miR-335 negatively regulates the characteristics of osteosarcoma stem cells by down regulation of POU5F1. MiR-335 combined with conventional chemotherapy drugs can synergistically inhibit osteosarcoma. |
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