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长链非编码RNA NEAT1的敲低通过抑制miR-194表达抑制骨肉瘤的增殖和侵袭并诱导其凋亡。

Knockdown of Long Non-Coding RNA NEAT1 Inhibits Proliferation and Invasion and Induces Apoptosis of Osteosarcoma by Inhibiting miR-194 Expression.

作者信息

Wang Heping, Yu Yanzhang, Fan Shuxin, Luo Leifeng

机构信息

Department of Orthopedics, Zhoukou Central Hospital, Zhoukou, China.

Department of Surgery, Zhoukou Central Hospital, Zhoukou, China.

出版信息

Yonsei Med J. 2017 Nov;58(6):1092-1100. doi: 10.3349/ymj.2017.58.6.1092.

Abstract

PURPOSE

Long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) has been implicated as an oncogene in the development and progression of osteosarcoma. This study aims to explore the mechanism of NEAT1 in osteosarcoma.

MATERIALS AND METHODS

Expressions of NEAT1 and miR-194 in osteosarcoma tissues and cells were detected by quantitative real-time PCR. The effects of NEAT1 knockdown or miR-194 overexpression on cell proliferation, invasion, and apoptosis were determined by 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyl tetrazolium bromide (MTT) assay, transwell invasive assay, and flow cytometry analysis, respectively. Luciferase reporter assay was performed to observe the possible interaction between NEAT1 and miR-194.

RESULTS

NEAT1 was upregulated and miR-194 was downregulated in osteosarcoma tissues and cells. Knockdown of NEAT1 or overexpression of miR-194 suppressed proliferation and invasion and induced apoptosis of osteosarcoma cells in vitro. Luciferase reporter assay validated that NEAT1 could interact with miR-194 and negatively modulated its expression. Furthermore, inhibition of miR-194 reversed the suppression of proliferation and invasion and the promotion of apoptosis induced by NEAT1 depletion in osteosarcoma cells.

CONCLUSION

Knockdown of NEAT1 suppressed proliferation and invasion and induced apoptosis in osteosarcoma cells by inhibiting miR-194 expression.

摘要

目的

长链非编码RNA(lncRNA)核旁斑组装转录本1(NEAT1)被认为是骨肉瘤发生发展过程中的一种癌基因。本研究旨在探讨NEAT1在骨肉瘤中的作用机制。

材料与方法

采用定量实时PCR检测骨肉瘤组织和细胞中NEAT1和miR-194的表达。分别通过3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐(MTT)法、Transwell侵袭实验和流式细胞术分析,检测NEAT1敲低或miR-194过表达对细胞增殖、侵袭和凋亡的影响。进行荧光素酶报告基因实验,观察NEAT1与miR-194之间可能的相互作用。

结果

骨肉瘤组织和细胞中NEAT1表达上调,miR-194表达下调。敲低NEAT1或过表达miR-194可抑制骨肉瘤细胞的体外增殖和侵袭,并诱导其凋亡。荧光素酶报告基因实验证实NEAT1可与miR-194相互作用,并对其表达起负调控作用。此外,抑制miR-194可逆转NEAT1缺失对骨肉瘤细胞增殖和侵袭的抑制作用以及对凋亡的促进作用。

结论

敲低NEAT1可通过抑制miR-194表达来抑制骨肉瘤细胞的增殖和侵袭,并诱导其凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ebf3/5653473/d830702feb33/ymj-58-1092-g001.jpg

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