Department of Orthopedic, The 117 Hospital of The PLA, Hangzhou, Zhejiang, 310012, China.
Department of Orthopedic, The 117 Hospital of The PLA, Hangzhou, Zhejiang, 310012, China.
Biomed Pharmacother. 2018 Aug;104:110-118. doi: 10.1016/j.biopha.2018.04.193. Epub 2018 May 15.
Osteosarcoma is a prevalent primary malignant tumor and long non-coding RNAs (lncRNAs) have been validated to modulate the osteosarcoma tumorigenesis. In present study, our research team investigates the role of a novel identified lncRNA DICER1-AS1 on the tumor progression and autophagy. Results showed that lncRNA DICER1-AS1 was up-regulated in osteosarcoma cells using microarray analysis and RT-PCR. Cellular functional experiments revealed that DICER1-AS1 knockdown suppressed the proliferation, migration, invasion and autophagy of osteosarcoma cells in vitro. Besides, DICER1-AS1 knockdown inhibited the protein expression levels of ATG5, LC3-II and Beclin 1, suggesting the inhibition on the autophagy of osteosarcoma cells. Moreover, miR-30b was verified to target 3'-UTR of DICER1-AS1 and ATG5 using bioinformatics tools and luciferase reporter assay or RNA-immunoprecipitation (RIP). Western blot showed that ATG5 protein expression was decreased in DICER1-AS1 knockdown and miR-30b mimics transfected cells, while increased in miR-30b inhibitor transfected cells, presenting a negative correlation with miR-30b and a positive correlation with DICER1-AS1. Finally, xenograft assay in vivo indicated that DICER1-AS1 knockdown inhibited the osteosarcoma tumor growth and protein expression level of ATG5. In summary, all the results conclude that DICER1-AS1 regulates the proliferation, invasion and autophagy of osteosarcoma via miR-30b/ATG5 axis, providing a novel insight for osteosarcoma tumorigenesis.
骨肉瘤是一种常见的原发性恶性肿瘤,长链非编码 RNA(lncRNA)已被证实可调节骨肉瘤的肿瘤发生。在本研究中,我们的研究团队研究了一种新鉴定的 lncRNA DICER1-AS1 对肿瘤进展和自噬的作用。结果显示,通过微阵列分析和 RT-PCR 显示骨肉瘤细胞中的 lncRNA DICER1-AS1 上调。细胞功能实验表明,DICER1-AS1 敲低抑制骨肉瘤细胞在体外的增殖、迁移、侵袭和自噬。此外,DICER1-AS1 敲低抑制骨肉瘤细胞中 ATG5、LC3-II 和 Beclin 1 的蛋白表达水平,表明对骨肉瘤细胞自噬的抑制作用。此外,通过生物信息学工具和荧光素酶报告基因检测或 RNA 免疫沉淀(RIP)验证 miR-30b 靶向 DICER1-AS1 和 ATG5 的 3'-UTR。Western blot 显示,DICER1-AS1 敲低和 miR-30b 模拟转染细胞中 ATG5 蛋白表达降低,而 miR-30b 抑制剂转染细胞中 ATG5 蛋白表达增加,与 miR-30b 呈负相关,与 DICER1-AS1 呈正相关。最后,体内异种移植实验表明 DICER1-AS1 敲低抑制骨肉瘤肿瘤生长和 ATG5 蛋白表达水平。总之,所有结果表明 DICER1-AS1 通过 miR-30b/ATG5 轴调节骨肉瘤的增殖、侵袭和自噬,为骨肉瘤的发生提供了新的见解。