LncRNA DICER1-AS1 通过 miR-30b/ATG5 促进骨肉瘤细胞的增殖、侵袭和自噬。

LncRNA DICER1-AS1 promotes the proliferation, invasion and autophagy of osteosarcoma cells via miR-30b/ATG5.

机构信息

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.

出版信息

Biomed Pharmacother. 2018 Aug;104:110-118. doi: 10.1016/j.biopha.2018.04.193. Epub 2018 May 15.

Abstract

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.

摘要

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。

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